Genomic DNA Ex trac tion and Purification Genomic DNA M ini K it (Blood/Cultured Cell)...30 Genomic DNA Maxi K it (Blood/Cultured Cell)...31 Genomic DNA M ini K it ( Tissue)...32 gsync TM DNA Ex trac tion K it...33 Genomic DNA M ini K it (Plant)...34 Genomic DNA Maxi K it (Plant)...35 Geneaid TM DNA Isolation K its...36 GENEzol TM DNA Reagent Plant...37 Presto TM M ini gdna Yeast K it...38 Presto TM M ini gdna Bac teria K it...39 Geneius TM M icro gdna Ex trac tion K it...40 Presto TM Buccal Swab gdna Ex trac tion K it...41 Presto TM DNA/RNA/Protein Ex trac tion K it...42 Presto TM 96 Well Blood gdna Ex trac tion K it...43 Presto TM 96 Well Plant gdna Ex trac tion K it...44 Presto TM 96 Well gdna Binding Plate...45
Genomic DNA M ini K it (Blood/Cultured Cell) Introduction (GB100, GB300) The Genomic DNA Mini Kit (Blood/Cultured Cell) is optimized for genomic, mitochondrial and virus DNA purification from whole blood, buffy coat and cultured cell samples. RBC Lysis Buffer and chaotropic salt are used to lyse cells and degrade protein, allowing DNA to bind to the glass fiber matrix of the spin column. Contaminants are removed using a Wash Buffer (containing ethanol) and the purified genomic DNA is eluted by a low salt Elution Buffer, TE or water. Phenol/chloroform extraction or alcohol precipitation is not required and the entire procedure can be completed within 25 mintues. Purify genomic DNA within 25 minutes! High Yield, High Quality Genomic DNA: 4-10 μg of pure genomic DNA (A260/A280 = 1.8-2.0) from 300 µl of whole blood Variety of samples: up to 300 µl of whole blood, up to 200 µl of buffy coat cultured cells (up to 1 x 10 7 ), yeast (up to 2 x 10 8 ) Elution volume: 30-200 µl Genomic DNA glass fiber spin columns (membrane optimized for genomic DNA extraction) Genomic DNA Mini Kit (Blood/Cell) Protocol Procedure PCR, AFLP/PADP, RFLP, Southern Blotting, Real-time PCR RBC Lysis Buffer GT Buffer GB Buffer Elution Buffer (10 mm Tris-HCl, ph8.5 at 25 º C) GD Columns 2 ml Collection Tubes RBC lysis of whole blood, buffy coat and cultured cell samples Cell lysis of blood, buffy coat and cultured cell samples Genomic DNA Mini Kit (Blood/Cultured Cell) Test Data Figure 1. Genomic DNA (20-30 kb) was extracted from 300 µl of whole blood using the Genomic DNA Mini Kit (Blood/Cultured Cell). The purified DNA was eluted in 200 µl of Elution Buffer and 15 µl aliquots of the final sample were analyzed by electrophoresis on a 1% agarose gel. M = Geneaid 1 Kb DNA Ladder Sample Yield 260/280 1. 300 µl 8.07 µg 1.87 2. 300 µl 8.18 µg 1.87 3. 300 µl 8.19 µg 1.87 Wash (removal of contaminants while DNA remains bound to membrane) M 1 2 3 Figure 2. Geneaid GD Columns are specially designed and pretreated to ensure high genomic DNA yields from a variety of samples. Page 30
Genomic DNA Maxi K it (Blood/Cultured Cell) Introduction (GDM010, GDM025) The Genomic DNA Maxi Kit (Blood/Cultured Cell) is optimized for DNA purification (including genomic, mitochondrial and viral DNA) from up to 10 ml of frozen or fresh blood and cultured cells. Protease is used to reduce hemoglobin contamination while chaotropic salt and detergents are used to lyse cells and degrade protein, allowing the DNA to bind to the glass fiber matrix of the spin column. Contaminants are removed using a Wash Buffer (containing ethanol) and the purified genomic DNA is eluted by a low salt Elution Buffer, TE or water. The entire procedure can be completed in 1 hour without phenol/chloroform extraction or alcohol precipitation. Purified DNA, with approximately 20-30 kb, is suitable for use in PCR or other enzymatic. Large Sample Size: up to 10 ml of frozen blood or fresh blood, cultured cells (up to 1 x 10 8 ) High Yield: up to 200 µg of pure genomic DNA from 10 ml of whole blood Glass fiber maxi spin column Elution volume: 500 µl-2 ml Genomic DNA Maxi Kit (Blood/Cell) Protocol Procedure PCR, AFLP/PADP, RFLP, Southern Blotting, Real-time PCR GB Buffer Elution Buffer (10 mm Tris-HCl, ph8.5 at 25 º C) Protease GD Maxi Columns Collection Tubes Genomic DNA Maxi Kit (Blood/Cultured Cell) Test Data M 1 2 3 Figure 1. Genomic DNA from 10 ml whole blood samples was extracted using the Genomic DNA Maxi Kit (Blood/Cultured Cell). 0.5 μl aliquots from 1 ml eluates of purified genomic DNA were analyzed by electrophoresis on a 0.8% agarose gel. M = Geneaid 1 Kb DNA Ladder Sample DNA Conc. 260/280 260/230 Yield 1 180.6 ng/µl 1.84 2.28 180.6 µg 2 195.8 ng/µl 1.86 2.15 195.8 µg 3 187.5 ng/µl 1.84 2.21 187.5 µg Cell lysis of blood and cultured cell samples Wash (removal of contaminants while DNA remains bound to membrane) Figure 2. Geneaid GD Maxi Columns are specially designed to ensure high genomic DNA yields from blood and cultured cell samples. Page 31
Genomic DNA M ini K it ( Tissue) Introduction (GT050, GT100, GT300) The Genomic DNA Mini Kit (Tissue) is optimized for genomic, mitochondrial and virus DNA purification from a variety of animal tissue and insects. This genomic DNA extraction kit includes DNase and RNase-free Micropestles to ensure efficient homogenization of tissue samples and reduce the time required for cell lysis. Proteinase K and chaotropic salt are used to lyse cells and degrade protein, allowing DNA to be easily bound by the glass fiber matrix of the spin column. Once any contaminants have been removed, using a Wash Buffer (containing ethanol), the purified DNA is eluted by a low salt Elution Buffer, TE or water and is ready for use in a variety of downstream Purify genomic DNA in 30 minutes! High yield: 10-20 µg (0.5 cm mouse tail), 20-50 µg (20 mg mouse kidney) Sample size: up to 30 mg of tissue (tailsnips, liver, kidney, brain, adipose tissue, earpunches, insects etc.) Genomic DNA glass fiber spin columns (membrane optimized for genomic DNA extraction) Elution volume: 30-200 µl Provided Micropestles are certified DNase and RNase-free Genomic DNA Mini Kit (Tissue) Protocol Procedure PCR, AFLP/PADP, RFLP, Southern Blotting, Real-time PCR Micropestle GT Buffer GBT Buffer Elution Buffer (10 mm Tris-HCl, ph8.5 at 25 º C) Proteinase K GS Columns 2 ml Collection Tubes Tissue dissociation in a 1.5 ml microcentrifuge tube using the provided micropestle Cell lysis of tissue samples Genomic DNA Mini Kit (Tissue) Test Data 1 2 3 4 M Figure 1. Genomic DNA was extracted using the Genomic DNA Mini Kit (Tissue). The purified genomic DNA (20-30 kb) was EcoRI digested and analyzed by electrophoresis on a 1% agarose gel. 1 = Mouse Liver, 2 = Mouse Tail, 3 = Fish Muscle, 4 = Fruit Fly (Drosophila), M = Geneaid 1 Kb DNA Ladder Mouse Tissue Tissue Size Yield Tail 0.5 cm 10-20 µg Liver 20 mg 10-20 µg Brain 20 mg 10-20 µg Lung 20 mg 5-10 µg Muscle 20 mg 5-10 µg Kidney 20 mg 10-20 µg Wash (removal of contaminants while DNA remains bound to membrane) Figure 2. Geneaid GD Columns are specially designed and pretreated to ensure high genomic DNA yields from tissue and insect samples. Page 32
gsync TM DNA Ex trac tion K it Introduction (GS050, GS100, GS300) The gsync DNA Extraction Kit is optimized for genomic, mitochondrial and virus DNA purification from whole blood (fresh blood and frozen blood), serum, plasma, buffy coat, body fluids, cultured cells, tissue, rodent tails, ear punches, formalin-fixed paraffin-embedded tissue (FFPE), amniotic fluid, hair, insects and sperm in one convenient kit. This DNA extraction kit uses Proteinase K and chaotropic salt to lyse cells and degrade protein, allowing DNA to bind to the glass fiber matrix of the spin column. Contaminants are removed using a Wash Buffer and the purified genomic DNA is eluted by a low salt Elution Buffer, TE or water. The entire procedure can be completed in as little as 20 minutes without phenol/chloroform extraction or alcohol precipitation. The purified DNA (approximately 20-30 kb) is suitable for use in PCR or other enzymatic. Purify genomic DNA in as little as 20 minutes! Variety of samples: 50-200 μl of whole blood, plasma, serum, buffy coat, up to 25 mg of tissue, rodent tails, ear punches, insects, up to 1 x 10 7 cultured cells, up to 25 mg of formalin-fixed-paraffin-embedded tissue, up to 15 ml of amniotic fluid, hair, up to 100 µl of sperm High yield: up to 5 µg from whole blood (260/280 = 1.8-2.0) NO manual homogenization of tissue samples Elution volume: 30-200 µl Genomic DNA glass fiber spin columns (membrane optimized for genomic DNA extraction) gsync DNA Extraction Kit Protocol Procedure Cell lysis of tissue samples PCR, AFLP/PADP, RFLP, Southern Blotting, Real-time PCR GST Buffer GSB Buffer Elution Buffer (10 mm Tris-HCl, ph8.5 at 25 º C) Proteinase K GS Columns 2 ml Collection Tubes The gsync DNA Extraction Kit Test Data Wash (removal of contaminants while DNA remains bound to membrane) Figure 1. Genomic DNA from 50, 100 and 200 µl whole blood samples was extracted using the gsync DNA Extraction Kit. 10 µl from 100 µl eluates of purified genomic DNA was analyzed by electrophoresis on a 0.8% agarose gel. 1-3 = 50 µl whole blood sample 4-6 = 100 µl whole blood sample 7-9 = 200 µl whole blood sample M = Geneaid 1 Kb DNA Ladder Volume Yield 260/280 50 µl 1.54 µg 1.85 100 µl 2.70 µg 1.87 200 µl 5.56 µg 1.90 Figure 2. Geneaid GD Columns are specially designed and pretreated to ensure high genomic DNA yields from a variety of samples. M 1 2 3 4 5 6 7 8 9 Page 33
Genomic DNA M ini K it (Plant) Introduction (GP100) The Genomic DNA Mini Kit (Plant) is optimzed for genomic, mitochondrial and chloroplast DNA purification from a variety of plant species. Plant samples are initially ground to a fine powder, followed by lysis buffer incubation. The lysate is treated with RNase A then filtered to remove cell debris and salt precipitates. In the presence of the binding buffer, coupled with chaotropic salt, genomic DNA in the lysate binds to the glass fiber matrix of the spin column. Contaminants are removed using a Wash Buffer (containing ethanol) and the purified genomic DNA is eluted by a low salt Elution Buffer, TE or water. DNA phenol extraction or alcohol precipitation is not required, and the entire procedure can be completed in as little as 25 minutes. Purify genomic DNA in as little as 25 minutes! High yield: 3-5 μg (100 mg Thale cress), 20-25 µg (100 mg Tobacco) Sample size: up to 100 mg (fresh plant tissue), up to 25 mg (dry plant tissue) Genomic glass fiber spin column (membrane optimized for genomic DNA purification from plant samples) Elution volume: 30-200 µl Genomic DNA Mini Kit (Plant) Protocol Procedure PCR, Real-time PCR, Southern Blotting and RFLP GPX1 Buffer (optimized for samples with high polysaccharide content) GP1, GP2, GP3 Buffers Elution Buffer (10 mm Tris-HCl, ph8.5 at 25 º C) Filter Columns GD Columns 2 ml Collection Tubes Cell lysis of plant samples Genomic DNA Mini Kit (Plant) Test Data Figure 1. Genomic DNA was extracted from a variety of 100 mg plant species using the Genomic DNA Mini Kit (Plant). A 3 µl aliquot from each 200 µl eluate was analyzed by electrophoresis on a 1% agarose gel. M = Geneaid 1 Kb DNA ladder 1. Cinnamomun camphora (Camphor tree) 2. Pisum sativum (Pea sprout) 3. Arabidopsis thaliana (Thale cress) 4. Oryza sativa (Rice) 5. Ipomoea batatas (Sweet potato vine) 6. Rhizoma dioscoreae (Chinese yam ) 7. Populus tremula (Aspen) 8. Flammulina velutipes (Mushroom) 9. Oxalis comiculats (Clover) Wash (removal of contaminants while DNA remains bound to membrane) M 1 2 3 4 5 6 7 8 9 Plant Species DNA Yield Thale Cress (Arabidopsis thaliana) 3~5 µg Rice (Oryza sativa) 10~15 µg Tomato (Lycopersicon esculentum) 10~15 µg Tobacco (Nicotiana tabacum) 20~25 µg Chinese Yam (Rhizoma dioscoreae) 30~60 µg Corn (Zea mays) 15~20 µg Sweet Potato (lpomoea batatas) 20~30 µg Orchid (Phalaenopsis aphrodite) 5~10 µg Camphor (Cinnamommun camphora) 15~20 µg Spinach (Spinacia oleracea) 5~10 µg Bamboo (Bambusa iodhamii) 10~15 µg Table 1. Genomic DNA yield of various plant species following DNA purification using the Genomic DNA Mini Kit (Plant). Figure 2. Geneaid GD Columns are specially designed and pretreated to ensure high genomic DNA yields from a variety of plant species. Page 34
Genomic DNA Maxi K it (Plant) Introduction (GPM10, GPM25) The Genomic DNA Maxi Kit (Plant) is optimized for DNA extraction, including genomic, mitochondrial and chloroplast DNA purification from plant tissue and cells using an efficient maxiprep system. Samples are disrupted by both grinding in liquid nitrogen and lysis buffer incubation. The lysate is treated with RNase A then filtered to remove cell debris and salt precipitates. In the presence of the binding buffer, coupled with chaotropic salt, the genomic DNA in the lysate binds to the glass fiber matrix of the spin column. Contaminants are removed using a Wash Buffer (containing ethanol) and the purified genomic DNA is eluted by a low salt Elution Buffer, TE or water. The procedure does not require DNA phenol extraction or alcohol precipitation, and can be completed in less than 1 hour. The purified genomic DNA is ready for use in PCR, Real-time PCR, Southern Blotting and RFLP. Binding Capacity: 500 μg Large sample size: up to 1 g (fresh plant tissue), up to 250 mg (dry plant tissue) Genomic glass fiber maxi spin column (membrane optimized for genomic DNA purification) Elution volume: 200 µl-2 ml PCR, Real-time PCR, Southern Blotting, RFLP GPX1 Buffer (optimized for samples with high polysaccharide content) GP1, GP2, GP3 Buffers Elution Buffer (10 mm Tris-HCl, ph8.5 at 25 º C) Filter Columns GD Maxi Columns Collection Tubes Genomic DNA Maxi Kit (Plant) Test Data Figure 1. Genomic DNA (approximately 30 kb) was extracted using the Genomic DNA Maxi Kit (Plant). 0.5 g of fresh lemon leaves and 0.5 g of fresh coffee leaves were ground into a fine powder followed by DNA extraction. 2 µl aliquots of purified genomic DNA from 1 ml eluates were Genomic DNA Maxi Kit (Plant) Protocol Procedure Cell lysis of plant samples Wash (removal of contaminants while DNA remains bound to membrane) M 1 2 3 4 Sample DNA Conc. 260/280 Yield Lemon Leaf 80.6 ng/µl 1.81 80.6 µg Lemon Leaf 82.2 ng/µl 1.80 82.2 µg Coffee Leaf 37.3 ng/µl 1.79 37.3 µg Coffee Leaf 36.4 ng/µl 1.75 36.4 µg Figure 2. Geneaid GD Maxi Columns are specially designed to ensure high genomic DNA yields from a variety of plant species. Page 35
Geneaid TM DNA Isolation K its Introduction Geneaid DNA Isolation Kits offer a simple and gentle reagent DNA precipitation method for isolating high molecular weight genomic, mitochondrial or viral DNA suitable for archiving or sensitive downstream applications. This highly versatile solution based system can be scaled proportionately in order to satisfy larger sample volumes, providing a convenient sample-storage procedure with minimal hands on time. Isolate high molecular weight DNA for archiving or sensitive downstream applications High yield, High quality DNA (A260/A280 = 1.8-2.0) Sample: 300 µl of whole blood, 3 ml of whole blood, 10 ml of whole blood, wide variety of bacteria strains: Gram (+) Positive and Gram (-) Negative bacterial cells (up to 1 x 10 9 ), cultured cells (up to 1 x 10 7 ), tissue RXNS: 100 and 1,000 Easily scalable to satisfy larger sample volumes Minimal hands on time Convenient, cost effective system similar to Gentra Puregene DNA archiving, PCR, AFLP, RFLP/PADP, Southern Blotting, Real-time PCR Geneaid DNA Isolation Kits Protocol Procedure Cell lysis of blood, bacteria, cultured cells and tissue Geneaid DNA Isolation Kit Blood (GEB100, GEB01K, GEB01K+) RBC Lysis Buffer, Cell Lysis Buffer, Protein Removal Buffer, RNase A, DNA Hydration Buffer Geneaid DNA Isolation Kit Bacteria (GEE300, GEE03K, GEE03K+) Gram+ Buffer, Lysozyme, Cell Lysis Buffer, Protein Removal Buffer, RNase A, DNA Hydration Buffer Geneaid DNA Isolation Kit Cultured Cell (GEC150, GEC1.5K, GEC1.5K+) Cell Lysis Buffer, Protein Removal Buffer, RNase A, DNA Hydration Buffer Geneaid DNA Isolation Kit Tissue (GET150, GET1.5K, GET1.5K+) Proteinase K, Cell Lysis Buffer, Protein Removal Buffer, RNase A, DNA Hydration Buffer Protein Removal DNA Precipitation Geneaid DNA Isolation Kit (Blood) Test Data Re-hydration of high molecular weight genomic DNA Figure 1. High molecular weight genomic DNA (approximately 100-200 kb) was extracted using both the Geneaid gdna Extraction Kit (Blood) and the Gentra Puregene Kit (Blood). Genomic DNA was extracted from 300 µl of whole human blood and resolved in 100 µl of DNA Hydration Buffer. 5 µl of the resolved genomic DNA was analyzed by electrophoresis on a 1% agarose gel. Lane 1-3 = Archive gdna Extraction Kit (Blood) Lane 4-6 = Gentra Puregene Kit (Blood) 1 2 3 4 5 6 Page 36
GENEzol TM DNA Reagent Plant Introduction (GR100, GR200) GENEzol DNA Reagent Plant provides a quick and easy 3 step CTAB and chloroform based method to isolate total DNA (including genomic, mitochondrial and chloroplast DNA) from a variety of plant species (including algae and cynobacteria). This unique reagent is able to lyse most common plant samples and plant samples with high polysaccharide content. Two procedures are available with this kit. The Fast PCR Grade DNA Protocol offers a simplified, time-saving method for DNA extraction. The purified DNA is suitable for routine PCR screening. Phenol and chloroform extraction is not required and can be completed in 50 minutes. The High Purity and Yield DNA Protocol offers a full procedure for extracting large amounts of pure DNA. The extracted total DNA is ready for use in PCR, Real-time PCR, Southern Blotting, Mapping and RFLP. The entire procedure can be completed in 80 minutes. High molecular weight genomic DNA extraction from a variety of plant species using a simple and gentle DNA precipitation method High yield, High quality DNA (A260/A280 = 1.8-2.0) Up to 1 g of fresh plant tissue and up to 0.5 g of dry plant tissue Convenient 3 step system using a single reagent Easily scalable to satisfy larger sample volumes Minimal hands on time Fast PCR Grade DNA protocol: 50 minutes High purity and high yield DNA protocol: 80 minutes Elution volume: 50-100 µl PCR, Real-time PCR, Southern Blotting, Mapping and RFLP GENEzol DNA Reagent Plant Protocol Procedure Cell lyis following plant sample preparation Phase separation GENEzol DNA Reagent Plant RNase A GENEzol DNA Reagent Plant Test Data Figure 1. Genomic DNA (approximately 30 kb) was extracted using GENEzol DNA Reagent Plant. 50 mg of fresh Arabidopsis leaves were initially ground in GENEzol DNA Reagent Plant. A 15 µl aliquot of extracted genomic DNA from a 100 µl eluate was analyzed by electrophoresis on a 1% agarose gel. M = Geneaid 1 Kb DNA Ladder DNA Precipitation Re-hydration of high molecular weight genomic DNA Test DNA Conc. 260/280 260/230 Yield 1 380.7 µg/ml 1.97 1.88 19.04 µg 2 316.9 µg/ml 1.96 1.89 15.85 µg M 1 2 Figure 2. GENEzol DNA Reagent Plant uses a simple CTAB and chloroform procedure to extract high-quality total DNA from a variety of plant species. Page 37
Presto TM M ini gdna Yeast K it Introduction (GBY100, GBY300) The Presto Mini gdna Yeast Kit offers a simple and gentle reagent DNA precipitation method for isolating high molecular weight genomic, mitochondrial or viral DNA from yeast and other fungus species. This highly versatile solution based system can be scaled proportionately in order to satisfy larger yeast sample sizes, providing a convenient procedure with minimal hands on time. The provided Sorbitol Buffer, when combined with β-mercaptoethanol, zymolase or lyticase, will efficiently lyse fungus cell walls consisting of chitin and polysaccharides. The extracted DNA (A260/A280 = 1.8-2.0) is suitable for use in PCR or other enzymatic. Extract genomic DNA from (up to 2 x 10 8 ) yeast and other fungus species using a simple and gentle 3 buffer system in 40 minutes A simple and gentle reagent DNA precipitation method Convenient premixed Sorbitol Buffer Elution volume: 50-100 µl Presto Mini gdna Yeast Kit Protocol Procedure PCR, AFLP, RFLP/PADP, Southern Blotting, Real-time PCR Sorbitol Buffer Cell Lysis Buffer Protein Removal Buffer DNA Hydration Buffer Presto Mini gdna Yeast Kit Test Data Figure 1. Genomic DNA (approximately 30 kb) was extracted using the Presto Mini gdna Yeast Kit. Saccharomyces cerevisiae (2 10 8 ) was harvested by centrifugation at 5,000 x g for 10 minutes. A 15 µl aliquot of extracted genomic DNA from a 100 µl eluate was analyzed by electrophoresis on a 1% agarose gel. M = Geneaid 1 Kb DNA Ladder Test DNA Conc. 260/280 260/230 Yield 1 115.5 µg/ml 1.91 1.87 11.6 µg 2 142.9 µg/ml 1.92 2.01 14.3 µg 3 137.1 µg/ml 1.91 1.97 13.7 µg Cell lysis of yeast and other fungus species Protein Removal DNA Precipitation M 1 2 3 Re-hydration of high molecular weight genomic DNA Figure 2. The Presto Mini gdna Yeast Kit uses a simple procedure to extract high-quality total DNA from yeast and other fungus species. Page 38
Presto TM M ini gdna Bac teria K it Introduction (GBB100, GBB300) The Presto Mini gdna Bacteria Kit is optimized for genomic and viral DNA purification from Gram (-) negative and Gram (+) positive bacterial cells. Gram+ Buffer, when combined with lysozyme, will efficiently lyse bacterial cell walls consisting of the peptidoglycan layer. Chaotropic salt is used to further lyse cells and degrade protein, allowing DNA to easily bind to the glass fiber matrix of the spin column. Contaminants are removed using a Wash Buffer (containing ethanol) and the purified genomic DNA is eluted by a low salt Elution Buffer, TE or water. Phenol/chloroform extraction or alcohol precipitation is not required. Purify DNA from a wide variety of both Gram+ and Gram- bacterial cell strains in as little as 20 minutes! High yield: up to 40 μg (1 x 10 9 Escherichia coli) 10-15 µg (1 x 10 9 Bacillus subtilis) High purity gdna: A260/A280 >1.9 (20-40 kb) Convenient: Includes Gram+ Buffer for preparing lysozyme solutions Genomic DNA glass fiber spin column (membrane optimized for gdna purification from bacteria and sterilised to remove bacteria contamination) Elution volume: 50-200 µl, Lysozyme is shipped at room temperature and should be stored at -20ºC for extended periods PCR, Restriction Endonuclease Digestions, Array, Southern Blots and various other enzymatic Gram+ Buffer GT, GB Buffer W1 and Wash Buffer Lysozyme Proteinase K Elution Buffer (10 mm Tris-HCl, ph8.5 at 25ºC) GD Columns 2 ml Collection Tubes Presto Mini gdna Bacteria Kit Test Data Figure 1. Genomic DNA (approx. 30 kb) was extracted using the Presto Mini gdna Bacteria Kit. An Escherichia coli (1 10 9 ) culture (OD600=2, 1 ml) was harvested by centrifugation at 16,000 x g for 1 minute. A 5 µl aliquot of purified genomic DNA from a 100 µl eluate was analyzed by electrophoresis on a 1% agarose gel. M = Geneaid 1 Kb DNA Ladder Presto Mini gdna Bacteria Kit Protocol Procedure Sample preparation and cell lysis of bacterial cells Wash (removal of contaminants while DNA remains bound to membrane) M 1 2 3 M 1 2 3 Test DNA Yield 260/280 260/230 1 38.16 µg 2.05 2.26 2 33.88 µg 2.05 2.27 3 39.02 µg 2.07 2.29 Figure 2. Genomic DNA (approximately 30 kb) was extracted using the Presto Mini gdna Bacteria Kit. A Bacillus subtilis (1 10 9 ) culture (OD600=2, 1 ml) was harvested by centrifugation at 16,000 x g for 1 minute. A 5 µl aliquot of purified genomic DNA from a 100 µl eluate was analyzed by electrophoresis on a 1% agarose gel. M = Geneaid 1 Kb DNA Ladder Test DNA Yield 260/280 260/230 1 12.28 µg 1.96 2.28 2 12.08 µg 1.97 2.30 3 12.56 µg 1.96 2.29 Figure 3. Geneaid GD Columns are specially designed and pretreated to ensure high genomic DNA yields from a variety of bacteria samples. Page 39
Geneius TM M icro gdna Ex trac tion K it Introduction (GMB100, GMB300) The Geneius Micro gdna Extraction Kit provides an efficient method for purifying DNA (including genomic and mitochondrial DNA) from small volumes of whole blood, dried blood spots and urine. Proteinase K and chaotropic salt are used to lyse cells and degrade protein, allowing DNA to bind to the glass fiber matrix of the spin column. Carrier RNA is included with this DNA extraction kit to improve the efficiency of DNA binding to the spin column membrane. Contaminants are removed using W1 Buffer and Wash Buffer (containing ethanol). The purified genomic DNA is eluted by a low salt Elution Buffer, TE or water. The entire procedure can be completed without phenol/chloroform extraction or alcohol precipitation, with an average DNA yield of 300 ng from 6 mm dried blood spots. Purified DNA, with approximately 20-30 kb, is suitable for use in PCR or other enzymatic. Purify 20-30 kb (A260/A280 = 1.8-2.0) genomic DNA from small volume blood samples, dried blood spots, urine and saliva in as little as 20 minutes! High yield: 300 ng of pure genomic DNA from 6 mm blood spots Genomic DNA glass fiber spin column (membrane optimized for genomic DNA purification from small volume samples) Elution volume: 30-100 µl PCR, Restriction Endonuclease Digestions, Array, Southern Blots and various other enzymatic S1 Buffer S2 Buffer Carrier RNA Elution Buffer (10 mm Tris-HCl, ph8.5 at 25 º C) Proteinase K GD Columns 2 ml Collection Tubes Geneius Micro gdna Extraction Kit Real-Time PCR Data Geneius Micro gdna Extraction Kit Protocol Procedure Sample preparation and cell lysis of blood, blood spots and urine samples Wash (removal of contaminants while DNA remains bound to membrane) Figure 2. Geneaid GD Columns are specially designed and pretreated to ensure high genomic DNA yields from a variety of samples. Figure 1. DNA was purified from blood samples (1 64 µl) using the Geneius Micro gdna Extraction Kit. DNA was eluted in 30 µl of Elution Buffer followed by a Real-time PCR assay performed with a 3 μl aliquot as template, primers (designed to amplify the ACTB gene), and Fast SYBR Green PCR Master Mix using the StepOnePlusTM Real-Time PCR system (Applied Biosystems). The results confirm the efficiency of DNA extraction from as low as 1 µl blood samples. Page 40
Presto TM Buccal Swab gdna Ex trac tion K it Introduction (GSK100, GSK300) The Presto Buccal Swab gdna Extraction Kit provides an efficient method for purifying DNA (including genomic, mitochondrial and viral DNA) from buccal cells. Proteinase K and chaotropic salt are used to lyse cells and degrade protein, allowing DNA to bind to the glass fiber matrix of the spin column. Carrier RNA is included with the kit to improve the efficiency of DNA binding to the spin column membrane. Contaminants are removed using W1 and Wash Buffer (containing ethanol). The purified genomic DNA is eluted by a low salt Elution Buffer, TE or water. Phenol/chloroform extraction or alcohol precipitation is not required. Purify genomic DNA from buccal swabs in 20 minutes! High yield: up to 2 μg of pure genomic DNA per buccal swab High Purity gdna: A260/A280 = 1.8-2.0 DNA Size: 20-30 kb Spin Columns: glass fiber membrane optimized for genomic DNA extraction Elution volume: 50-100 µl Presto Buccal Swab gdna Extraction Kit Protocol Procedure PCR, Restriction Endonuclease Digestions, Array, Southern Blots and various other enzymatic S1, S2 Buffer Carrier RNA Elution Buffer (10 mm Tris-HCl, ph8.5 at 25 º C) Proteinase K GD Columns, Filter Columns 2 ml Collection Tubes Sample collection Sample preparation and cell lysis Presto Buccal Swab gdna Kit Test Data Figure 1. Genomic DNA from 3 individual cotton buccal swab samples was extracted using the Presto Buccal Swab gdna Extraction Kit. The purified genomic DNA was analyzed by electrophoresis on a 0.8% agarose gel. M = Geneaid 1 Kb DNA Ladder M 1 2 3 Sample DNA Conc. 260/280 Yield 1 52.20 ng/µl 1.86 1.83 µg 2 34.80 ng/µl 1.90 1.22 µg 3 50.90 ng/µl 1.92 1.78 µg Wash (removal of contaminants while DNA remains bound to membrane) Figure 2. The purified genomic DNA was used as a template for amplifying partial human 5S ribosomal DNA (91 bp) by PCR. The PCR products were analyzed on a 0.8% agarose gel. M = Geneaid 1 Kb DNA Ladder M 1 2 3 Figure 3. Geneaid GD Columns are specially designed and pretreated to ensure high genomic DNA yields from buccal cell samples. Page 41
Presto TM DNA/RNA/Protein Ex trac tion K it Introduction (DRP050, DRP100) The PrestoTM DNA/RNA/Protein Extraction Kit provides an efficient method for purifying genomic DNA, total RNA and total protein simultaneously from a variety of samples (cultured cells, animal tissue, whole blood and biological liquids). Chaotropic salt is used to lyse cells and inactive DNases and RNases, allowing DNA to bind to the genomic DNA spin column. The flow-through can then be transferred to the RNA spin column for RNA binding. The proteins in the flow-through can then be precipitated using acetone. Contaminants are effectively removed using wash buffers followed by pure genomic DNA elution in a low salt buffer and pure total RNA elution in RNase-free Water. DNA/RNA purification can be completed in 25 minutes without phenol/chloroform extraction or alcohol precipitation and protein purification can be completed in 50 minutes. The purified DNA, with approximately 20-30 Kb, is suitable for use in PCR or other enzymatic and the purified RNA (including mirna) is ready for use in RT-PCR, Real-time PCR, northern blotting, primer extension, mrna selection and cdna synthesis. The purified proteins can be directly analyzed on a SDS-PAGE and subsequent western blot. Purify genomic DNA and total RNA in 25 minutes! High yield: up to 9 μg of genomic DNA, 20 μg of total RNA, 120 μg of protein from 1.5 x 106 HeLa cells Sample: cultured cells, animal tissue, whole blood, biological fluids Format: genomic DNA spin column and total RNA spin column certified DNase and RNase-free Elution volume: 50-200 μl (genomic DNA) / 25-50 μl (total RNA) Storage: dry at room temperature (15-25ºC) for up to 9 months Presto DNA/RNA/Protein Extraction Kit Protocol Procedure Sample preparation and Cell lysis DNA/RNA Separation: DNA binding and RNA flow-through The purified DNA, approximately 20-30 kb, is suitable for use in PCR or other enzymatic and the purified RNA (including mirna) is ready for use in RT-PCR, Real-time PCR, northern blotting, primer extension, mrna selection and cdna synthesis. RBC Lysis Buffer DR Buffer RW1 Buffer RPE Buffer RNase-free Water Elution Buffer (10 mm Tris-HCl, ph8.5 at 25ºC) GD Columns RB Columns 2 ml Collection Tubes DNA wash DNA elution Protein Precipitation on ice for 30 minutes Protein flow-through RNA binding RNA wash Protein Resuspension Presto DNA/RNA/Protein Kit Test Data Figure 1. Genomic DNA and Total RNA from 1 x 106 SSN-1 cells was extracted using the Presto DNA/RNA/Protein Extraction Kit. 10 µl from 100 µl eluates of purified genomic DNA and 10 µl from 50 µl eluates of purified total RNA was analyzed by electrophoresis on a 1% agarose gel. 1-2 = DNA from 1 x 106 SSN-1 cells 3-4 = RNA from 1 x 106 SSN-1 cells M = Geneaid 1 Kb DNA Ladder Sample 1. DNA 2. DNA 3. RNA 4. RNA M 1 2 3 µg/ml 48.3 46.7 432.8 426.7 260/280 1.92 1.92 2.11 2.11 260/230 2.10 1.99 2.24 2.22 Yield 4.83 µg 4.67 µg 21.64 µg 21.34 µg 4 RNA elution 100ºC for 3-5 minutes SDS-PAGE Analysis Figure 2. Geneaid GD Columns are specially designed and pretreated to ensure high genomic DNA yields from a variety of samples. Geneaid RB Columns are specially designed and pretreated to ensure high total RNA yields from a variety of samples. Page 42
Presto TM 96 Well Blood gdna Ex trac tion K it Introduction (96GBP04, 96GBP10) The Presto 96 Well Blood Genomic DNA Extraction Kit was designed for high-throughput purification of genomic, mitochondrial and virus DNA from whole blood (fresh blood and frozen blood), plasma, serum, body fluids, lymphocytes and cultured cells. This DNA extraction kit uses protease and chaotropic salt to lyse cells and degrade protein, allowing DNA to bind to the glass fiber matrix of the Presto gdna 96 Well Binding Plate. Contaminants are removed using a Wash Buffer and the purified genomic DNA is eluted by a low salt Elution Buffer, TE or water. The procedure can be completed within 45 minutes without phenol/chloroform extraction or alcohol precipitation. The purified DNA (approximately 20-30 kb) is suitable for use in PCR or other enzymatic. Purify 96 wells of genomic DNA in 45 minutes! High yield: up to 6 μg of genomic DNA from 200 μl of whole blood Sample: up to 200 μl of whole blood, plasma, serum and up to 5 x 106 lymphocytes or cultured cells per well Format: PrestoTM gdna 96 Well Binding Plates Elution volume: 200 μl Storage: dry at room temperature (15-25ºC) Presto 96 Well Blood gdna Extraction Kit Protocol Procedure Transfer up to 200 μl of whole blood, plasma, serum, body fluids, 5 x 106 lymphocytes, cultured cells to Microtubes (Racked) The purified DNA, approximately 20-30 kb, is suitable for use in PCR or other enzymatic DNA Binding GB Buffer Protease Elution Buffer (10 mm Tris-HCl, ph8.5 at 25ºC) PrestoTM gdna 96 Well Binding Plates Microtubes (Racked) Microtubes (8-strip) Caps for Microtubes (8-strip) 96 Deep Well Plates Wash Presto 96 Well Blood gdna Kit Test Data Figure 1. Genomic DNA was extracted from 200 μl whole human blood samples using the PrestoTM 96 Well Blood Genomic DNA Extraction Kit. The purified genomic DNA was eluted in 200 µl of Elution Buffer and 15 µl aliquots of the final sample (chosen from 4 random wells) were analyzed by electrophoresis on a 1% agarose gel. M = GeneaidTM 1 Kb DNA Ladder Sample 1. 200 µl blood 2. 200 µl blood 3. 200 µl blood 4. 200 µl blood M 1 2 3 ng/µl 27.6 28.2 32.9 25.9 260/280 1.81 1.75 1.74 1.81 Yield 4.7 µg 4.8 µg 5.6 µg 4.4 µg 4 Elution of pure DNA into Microtubes (Racked) Figure 2. PrestoTM 96 Well Blood Genomic DNA Extraction Kits can be used efficiently with either vacuum or centrifuge. Page 43
Presto TM 96 Well Plant gdna Ex trac tion K it Introduction (96GPP04, 96GPP10) The Presto 96 Well Plant Genomic DNA Extraction Kit is designed for high-throughput purification of total DNA (including genomic DNA, mitochondrial and chloroplast DNA) from various plant species. Homogenized samples are treated with RNase A then centrifuged to remove cell debris and salt precipitates. In the presence of a binding buffer, genomic DNA in the lysate binds to the glass fiber matrix of the Presto 96 Well gdna Binding Plate. Contaminants are removed using a Wash Buffer and the purified genomic DNA is eluted by a low salt Elution Buffer, TE or water. The entire procedure can be completed within 60 minutes without phenol/chloroform extraction or alcohol precipitation. The purified DNA (approximately 20-30 kb) is suitable for use in PCR or other enzymatic. 96 wells of pure genomic DNA from a variety of plants in 60 minutes! Sample Size: 100 mg (fresh plant tissue), 25 mg (dry plant tissue) per well Yield: up to 30 μg per well of genomic DNA from 100 mg of fresh plant tissue Format: Presto 96 Well gdna Binding Plates Elution volume: 200 μl Presto 96 Well Plant gdna Extraction Kit Protocol Procedure Centrifuge or vacuum manifold protocol Transfer plant samples to Microtubes (Racked) Storage: dry at room temperature (15-25ºC) for at least 1 year PCR, AFLP/PADP, RFLP, Southern Blotting, Real-time PCR GP1 Buffer, GPX1 Buffer GP2 Buffer, GP3 Buffer Elution Buffer (10 mm Tris-HCl, ph8.5 at 25ºC) RNase A PrestoTM gdna 96 Well Binding Plates Microtubes (Racked) Microtubes (8-strip) Caps for Microtubes (8-strip) 96 Deep Well Plates DNA Binding The Presto 96 Well Plant Kit Test Data Figure 1. Genomic DNA was extracted from 50 mg of fresh corn leaf samples using the PrestoTM 96 Well Plant Genomic DNA Extraction Kit. The purified genomic DNA was eluted in 200 µl of Elution Buffer and 15 µl aliquots of the final sample (chosen from 3 random wells) were analyzed by electrophoresis on a 1% agarose gel. M = GeneaidTM 1 Kb DNA Ladder Sample 1 2 3 M 1 2 DNA Conc. 77.7 ng/µl 78.8 ng/µl 76 ng/µl 260/280 1.80 1.80 1.79 260/230 2.02 2.05 1.99 Wash Yield 13.6 µg 13.8 µg 13.3 µg Elution of pure DNA into Microtubes (Racked) 3 Figure 2. PrestoTM 96 Well Plant Genomic DNA Extraction Kits can be used efficiently with either vacuum or centrifuge. Page 44
Presto gdna 96 Well Binding Plate Introduction (96GBP005, 96GBP100) The Presto gdna 96 Well Binding Plate provides high-throughput purification of genomic DNA from a variety of samples such as blood, serum, plasma, body fluids, up to 5 x 106 lymphocytes and cultured cells, corn leaf, mouse liver, mouse kidney, mouse tail snips etc. The glass fiber membrane is optimized for efficient DNA binding from a wide variety of sample types. The blue o-ring stabilizes the white membrane during high-speed centrifugation and the column tip ensures uninhibited flow-through while eliminating cross contamination of adjacent wells. With up to 6 µg of pure genomic DNA/well from 200 µl of blood, Presto gdna 96 Well Binding Plates are the high performance and cost effective solution for large scale genomic DNA extraction. High Efficiency: up to 6 µg of genomic DNA/well from 200 µl blood, 6-8 µg of genomic DNA/well from HeLa cells, 10-15 µg of genomic DNA/well from mouse liver and kidney, 10-20 µg of genomic DNA/well from mouse tails, 10-15 µg of genomic DNA/well from corn leaf Consistent high through-put recovery Presto 96 Well gdna Binding Plate Protocol Procedure The new column tip design ensures uninhibited flow-through while eliminating cross contamination of adjacent wells Glass fiber membrane optimized for genomic DNA extraction Blue o-ring stabilizes the glass fiber membrane during high-speed centrifugation and vacuum PCR, AFLP/PADP, RFLP, Southern Blotting, Real-time PCR DNA Binding GP1 Buffer, GPX1 Buffer GP2 Buffer, GP3 Buffer Elution Buffer (10 mm Tris-HCl, ph8.5 at 25ºC) RNase A PrestoTM gdna 96 Well Binding Plates Microtubes (Racked) Microtubes (8-strip) Caps for Microtubes (8-strip) 96 Deep Well Plates Wash 96 Well Column Specifications 1. Blue O-ring stabilizes the membrane during high-speed centrifugation 2. White membrane designed specifically for genomic DNA binding 3. Column tip ensures uninhibited flow-through while eliminating cross contamination of adjacent wells Elution of pure DNA into Microtubes (Racked) Figure 1. PrestoTM 96 Well gdna Binding Plates can be used efficiently with vacuum and centrifuge and 96 well instruments. Page 45