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GENET BIO DNA AMPLIFICATION PRODUCTS GUIDE Keynote of Products Prime TaqTM DNA Polymerase Prime TaqTM Premix ExPrime TaqTM DNA Polymerase ExPrime TaqTM Premix Ability Test of Products Comparative Test of Products Fidelity Test of Products
Keynote of Products Unique Compositions of eaction R Buffer Relax the secondary structure of template Improve the primer annealing Improve the PCR yield In case of using the general reaction buffer... Primer Denaturation/ Annealing A B M A B PCR In case of using the GeNet Bio reaction buffer... Denaturation/ Annealing Primer PCR M: DNA Marker Lane A: Company P Lane B: GeNet Bio PCR enzymes Keynote of Products 3
Prime Taq TM DNA Polymerase Prime Taq TM DNA Polymerase Feature General PCR enzyme Guarantees reproducible test result by its characteristic of high purity Amplifies target DNA by optimized buffer system Easily obtains under 5 Kb of DNA amplified products P rime Taq TM DNA Polymerase can be used in T-vector cloning Description: Prime Taq TM DNA Polymerase is a high quality recombinant enzyme and catalyze 5' 3' synthesis of DNA. The enzyme has no detectable 3' 5' proofreading exonuclease activity. It is provided with 0X reaction buffer that contains PCR enhancers. This reaction buffer will enable or improve sub-optimal PCR caused by templates that have a high degree of secondary structure or that are GC-rich. Cloning of PCR Products: A DNA fragment which is amplified by Prime Taq TM DNA Polymerase has A-overhang, and it enables you to do cloning by using T-vector. Quality Control: Endonuclease, Exonuclease, DNase, RNase and Protease activity is not detected. Prime Taq TM DNA Polymerase is determined to be 90% pure as judged by SDS -PAGE. Unit Definition: One unit is defined as the amount of enzyme required to catalyze the incorporation of 0 nmoles of dntps into acid-insoluble material in 30 minutes at 74. Buffer and Reagents: Storage Buffer: 20 mm Tris-HCl (ph 8.0), 00 mm KCl, 0.5 mm EDTA, 0. mm DTT, 0.5% Tween 20, 0.5 % Nonidet P-40, 50% Glycerol 0X Reaction Buffer: Contains Tris-HCl (ph 9.0), PCR enhancers, (NH4)2SO4, 20 mm MgCl2 0 mm dntps Mixture: 2.5 mm each of datp, dctp, dgtp and dttp Cat. No. A-type G-000 250 units G-00 250 units X 2 G-002 250 units X 4 B-type G-000-250 units G-00-250 units X 2 G-002-250 units X 4 Conc. of Enzyme 5 units/ Supplied with A-type 0X Reaction Buffer (with 20 mm MgCl2) with 0 mm dntps Mixture B-type 0X Reaction Buffer (with 20 mm MgCl2) without dntps Mixture Storage Store at -20 4
Prime Taq TM Premix Feature Contains mixture of rime P Taq TM DNA Polymerase, dntps, reaction buffer in 2X concentration Contains loading dye and sediment for immediate loading to gel after reaction P rovides the same result compare with manual formula P rime Taq TM Premix can be used in T-vector cloning Cat. No. G-2000 ml X G-200 ml X 3 G-2002 ml X 5 Description: Prime Taq TM Premix contains Prime Taq TM DNA Polymerase, reaction buffer, dntps mixture, protein stabilizer, and optimizes the convenience to use by adding sediment for electrophoresis and 2X solution of loading dye. In general, Prime Taq TM Premix shows no decline of activity compare with Prime Taq TM DNA Polymerase, even in a room temperature. Prime Taq TM Premix is good for under 5 Kb of PCR products, and also can obtain the result under 0 Kb by altering of some conditions (concentration of template, primer, and DNA Polymerase or increase of extension time). Cloning of PCR Products: A DNA fragment which is amplified by Prime Taq TM Premix has A-overhang, and it enables you to do cloning by using T-vector. Quality Control: Endonuclease, Exonuclease, DNase, RNase and Protease activity is not detected. Prime Taq TM DNA Polymerase is determined to be 90% pure as judged by SDS -PAGE. Unit Definition: One unit is defined as the amount of enzyme required to catalyze the incorporation of 0 nmoles of dntps into acid-insoluble material in 30 minutes at 74. Composition of 2X Premix Solution: Prime Taq TM DNA Polymerase unit/0 l, Tris-HCl (ph 9.0), PCR enhancer, (NH4)2SO4, 4 mm MgCl2, enzyme stabilizer, sediment, loading dye, 2.0 mm dntps mixture. Prime Taq TM Premix 5
ExPrime Taq TM DNA Polymerase ExPrime Taq TM DNA Polymerase Feature Has the ability of 3' 5' exonuclease function (High Fidelity) Has an effect for DNA amplification of cloning and sequencing purpose Easily obtains under 0 Kb of DNA amplified products (Long PCR) Obtains high PCR yield compare with Prime Taq TM DNA Polymerase ExPrime Taq TM DNA Polymerase can be used in T-vector cloning Description: ExPrime Taq TM DNA Polymerase has both 5' 3' exonuclease and 3' 5' exonucleas functions. Therefore, it has 3 times less error-rate compare with Taq DNA Polymerase, and it is efficient for cloning (High Fidelity). Also, it is easy to obtain PCR products in case of over 5 Kb as well as under 0 Kb of DNA amplified products (Long- PCR). Also ExPrime Taq TM DNA Poymerase gives you the satisfying result under 20 Kb by altering of some conditions (concentration of template, primer, DNA Polymerase or increase of extension time). Cloning of PCR Products: A DNA fragment which is amplified by ExPrime Taq TM DNA Polymerase has A-overhang, and it enables you to do cloning by using T-vector. Quality Control: Endonuclease, Exonuclease, DNase, RNase and Protease activity is not detected. ExPrime Taq TM DNA Polymerase is determined to be 90% pure as judged by SDS -PAGE. Unit Definition: One unit is defined as the amount of enzyme required to catalyze the incorporation of 0 nmoles of dntps into acid-insoluble material in 30 minutes at 74. Buffer and Reagents: Storage Buffer: 20 mm Tris-HCl (ph 8.0), 00 mm KCl, 0.5 mm EDTA, 0. mm DTT, 0.5% Tween 20, 0.5 % Nonidet P-40, 50% Glycerol 0X Reaction Buffer: Contains Tris-HCl (ph 9.0), PCR enhancers, (NH4)2SO4, 20 mm MgCl2 0 mm dntps Mixture: 2.5 mm each of datp, dctp, dgtp and dttp Cat. No. A-type G-4000 250 units G-400 250 units X 2 G-4002 250 units X 4 B-type G-4000-250 units G-400-250 units X 2 G-4002-250 units X 4 Conc. of Enzyme 5 units/ Supplied with A-type 0X Reaction Buffer (with 20 mm MgCl2) with 0 mm dntps Mixture B-type 0X Reaction Buffer (with 20 mm MgCl2) without dntps Mixture Storage Store at -20 6
ExPrime Taq TM Premix Feature Contains Mixture of ExP rime Taq TM DNA Polymerase, dntps, reaction buffer in 2X concentration Contains oading L dye and sediment for immediate loading to gel after reaction P rovides the same result compare with manual formula ExP rime Taq TM Premix can be used in T-vector cloning Cat. No. G-5000 ml X G-500 ml X 3 G-5002 ml X 5 Description: ExPrime Taq TM Premix is composed of ExPrime Taq TM DNA Polymerse, reaction buffer, dntp mixture, protein stabilizer and sediment which is needed for electrophoresis, and 2X solution type of loading dye, and these components maximize the convenience of the users. This product also has both 5 3 exonuclease and 3 5 exonucleas function that ExPrime Taq TM DNA Polymerase has. ExPrime Taq TM Premix shows no decline of activity compare with ExPrime Taq TM DNA Polymerase, even in a room temperature. Also, ExPrime Taq TM Premix gives you the satisfying result under 0 Kb as well as under 20 Kb by altering of some conditions (concentration of template, primer, DNA Polymerase or increase of extension time). Cloning of PCR Products: A DNA fragment which is amplified by ExPrime Taq TM Premix has A-overhang, and it enables you to do cloning by using T-vector. Quality Control: Endonuclease, Exonuclease, DNase, RNase and Protease activity is not detected. ExPrime Taq TM DNA Polymerase is determined to be 90% pure as judged by SDS -PAGE. Unit Definition: One unit is defined as the amount of enzyme required to catalyze the incorporation of 0 nmoles of dntps into acid-insoluble material in 30 minutes at 74. Composition of 2X Premix Solution: ExPrime Taq TM DNA Polymerase unit/0 l, Tris-HCl (ph 9.0), PCR enhancer, (NH4)2SO4, 4 mm MgCl2, enzyme stabilizer, sediment, loading dye, 2.0 mm dntps mixture. ExPrime Taq TM Premix 7
Ability Test of Products Working Range Test - Working range: Lambda DNA 0.5~20 Kb - PCR condition template: Lambda DNA 00 pg 94 5min 94 30sec, 58 30sec, 72 min 30 cycles 72 5min (5 Kb, 20 Kb size: 2 step PCR 94 30sec, 68 5min 30 cycles) Prime Taq TM DNA Polymerase M 2 3 4 5 6 7 8 9 M ExPrime Taq TM DNA Polymerase M 2 3 4 5 6 7 8 9 0 M2 Sequence Specific PCR Test - Target: Two polymorphisms of human IL-8 promoter site (position -607 and -37) -PCR products size Position -37: 446 bp, 26 bp Position -607: 30 bp, 96 bp Prime Taq TM DNA Polymerase ExPrime Taq TM DNA Polymerase -37-607 -37-607 a M b c M d a M b c M d Ability Test of Products M: Kb DNA Ladder, M2: Lambda DNA/Hind III Markers lane : 0.5 Kb, lane 2: Kb, lane 3: 2 Kb, lane 4: 3 Kb, lane 5: 4 Kb lane 6: 5 Kb, lane 7: 7 Kb, lane 8: 8 Kb, lane 9: 9 Kb, lane 0: 5 Kb lane :20 Kb M: 00 bp DNA Ladder a: Wild type (position -37), b: Hetero mutant (position -37) c: Homo mutant (position -607), d: Hetero mutant (position -607) 8
DNA Amplification Products Ability Test of Products HPV Detection Test Genomic DNA Contamination Test - Template: Extracted DNA from genetal sample - PCR condition 94 5min 94 30sec, 55 30sec, 72 30sec 35 cycles, 72 5min 94 5min 94 30sec, 55 30sec, 72 30sec 2 cycles, 72 5min Prime TaqTM DNA Polymerase M 2 3 4 st PCR 2nd PCR (nested) - This test is operated for the identification of E. coli genomic DNA contamination with positive (contain the E. coli genomic DNA) and negative (not contain the template) sample by PCR (30 ~ 50 cycles). M C 2 C 2 3 C 2 3 ExPrime TaqTM DNA Polymerase M 2 3 4 A B C Panel A: GeNet Bio Prime TaqTM DNA Polymerase Panel B: Company A standard Taq DNA Polymerase Panel C: Company P standard Taq DNA Polymerase M: 00 bp DNA Ladder, C: control (positive) lane : 30 cycles, lane 2: 40 cycles, lane 3: 50 cycles GENET BIOㆍwww.genetbio.com Ability Test of Products M: 00 bp DNA Ladder lane : Negative lane 2: HPV type 6 lane 3: HPV type 3 lane 4: HPV type 35 3 9
Comparative Test of Products PCR est T from Hela cdna Library M 2 3 4 5 M PCR est T from Fungi Genomic DNA M 2 3 4 5 6 M M: Kb DNA Ladder Lane : Prime Taq TM DNA Polymerase Lane 2: Prime Taq TM Premix Lane 3: ExPrime Taq TM DNA Polymerase Lane 4: ExPrime Taq TM Premix Lane 5: Company P M: Kb DNA Ladder Lane : Prime Taq TM DNA Polymerase Lane 2: Prime Taq TM Premix Lane 3: ExPrime Taq TM DNA Polymerase Lane 4: ExPrime Taq TM Premix Lane 5: Company P Lane 6: Company T Comparative Test of Products HB V Detection Test from Blood, Saliva and Breast Milk Blood Saliva Breast milk M 2 3 4 5 M 2 3 4 5 M 2 3 4 5 M M: Kb DNA Ladder Lane : Prime Taq TM DNA Polymerase Lane 2: Prime Taq TM Premix Lane 3: ExPrime Taq TM DNA Polymerase Lane 4: ExPrime Taq TM Premix Lane 5: Company P 0
Fidelity Test of Products F idelity of Prime Taq TM DNA Polymerase & ExPrime Taq TM DNA Polymerase Relative Error Frquency.2 Enzyme LacZ + (blue colonies) LacZ- (white colonies) % Mutant Errors Rate Relative Error Frequency Prime Taq TM DNA Polymerase ExPrime Taq TM DNA Polymerase Pfu DNA Polymerase 3,090 2,527 2,42,068 30 64 25.7 0.6 2.6 7.34 X 0-5 3.04 X 0-5 0.73 X 0-5 0.4 0. Relative Error Frequency 0.8 0.6 0.4 0.2 0.3 0. 0 Prime Taq TM DNA Pol. ExPrime Taq TM DNA Pol. * Reference. Wayne M. Barnes The fidelity of Taq DNA Polymerase catalyzing PCR is improved by an N-terminal deletion. GENE 2(992). 29-35 2. Wayne M. Barnes PCR amplification of up to 35-Kb DNA with high fidelity and high yield from lambda bacteriophage templates. Proc. Natl. Acad. Sci. USA 9(994). 226-2220 Enzyme Pfu DNA Pol. Fidelity Test of Products
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