10x Hot-Taq Buffer 1 ml 1 ml x 2ea 1 ml x 10ea. dntp Mix (each 10 mm) None / 0.2 ml None / 0.4 ml None / 0.4 ml x 5ea

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1 HelixAmp TM Hot-Taq Polymerase (Ver. 2.0) CERTIFICATE OF ANALYSIS (1603-V01R03) Contents HelixAmp TM Hot-Taq Polymerase (Ver. 2.0) Cat. No. HT250/HT250N HT500/HT500N HT2500/HT2500N Hot-Taq (2.5 units/μl) 0.1 ml 0.2 ml 0.2 ml x 5ea 10x Hot-Taq Buffer 1 ml 1 ml x 2ea 1 ml x 10ea dntp Mix (each 10 mm) None / 0.2 ml None / 0.4 ml None / 0.4 ml x 5ea 5x TuneUp TM solution None / 0.5 ml None / 0.5 ml x 2ea None / 0.5 ml x 10ea 6x Loading Dye 0.5 ml 1 ml 1 ml x 5ea Blue Box 1ea Certificate Analysis 1 ea 1 ea 1ea Description HelixAmp Hot-Taq polymerase (Ver. 2.0) is a chemically-modified form of purified Taq DNA polymerase and quite suitable for high-specific hot-start PCR, real-time PCR and multiplex PCR. The attached heat-labile chemical moiety makes Taq DNA polymerase inactive and suppresses the polymerization from nonspecifically bound primers which occurs during the setting of PCR mix and first ramp-up of thermal cycling. During the first denaturation step of PCR the chemical moieties are released from Taq polymerase and the enzyme turns to be active. With the high specificity HelixAmp Hot-Taq polymerase (Ver. 2.0) shows high performance in the reactions of genotyping (microsatellite or SNP), mutiplex PCR, and realtime PCR. For the maximum performance extremely pure dntps and TuneUp solutions are also included. TuneUp solution helps the DNA polymerase to efficiently amplify the problematic target region of high G+C content or structural problem. Application Storage buffer Hot-Start PCR 50% Glycerol, 20 mm Tris-HCl (ph 8.0), Real-Time PCR 100 mm KCl, 0.1 mm EDTA, 1 mm DTT, Genotyping 0.5% Tween 20, 0.5% Nonidet P-40 Multiplex PCR Concentration 2.5 units/μl Store -20

2 Hot-Taq Polymerase (Ver. 2.0) Quality Control Assay Contamination Assay for endo- and exodeoxyribonuclease HelixAmp Hot-Taq polymerase (Ver. 2.0) is a chemical-modified form of HelixAmp Taq polymerase. HelixAmp Taq polymerase was passed from quality control assay for contamination of endo- or exodeoxyribonuclease and the bacterial host DNA host using genomic sequencespecific primer set from host bacteria. Quality authorized by Youn Taek Go Protocol Although precipitates could be arised in the 10x Buffer, they will not affect the enzyme activities 1. Recommended amount of template DNA Human genomic DNA : 10 ~ 100 ng Bacterial genomic DNA : 5 ~ 50 ng Purified plasmid or phage DNA : 1 ~ 5 ng 2. Mix following components in a PCR tube Components Template 10x Hot-Taq Buffer dntp Mix (each 10 mm) Forward Primer (10 pmoles/μl) Reverse Primer (10 pmoles/μl) 5x TuneUp solution Distilled water Volumes (μl) X μl 5 μl 1 μl 0 ~ 20 μl 1.25 units to 50 μl TuneUp Solution is an additive altering the binding behavior of primer and template and can help the amplification that do not work well under standard PCR condition. Especially, TuneUp Solution can be used for the amplification of problematic template, such as high G+C content and repeat sequence regions. TuneUp Solution uses as adding into PCR reaction mixture from 0.5x to 2x.

3 Hot-Taq Polymerase (Ver. 2.0) 3. PCR condition Temperature & time Cycles 95, 15 min x 1 95, 20 sec Annealing Temp., 40 sec x 25 ~ 40 72, 1 min/kb (Expected size of product) 72, 5 min x 1 Annealing Temp. = T m (6 ~ 8 ) T m (Melting Temp.) = [4 x (G + C)] + [2 x (A + T)] Products Cat. No. Products Size HT250 HT250N HT500 HT500N HT2500 HT2500N (with 10x Hot-Taq Buffer) (with 10x Hot-Taq Buffer, 5x TuneUp solution, dntp Mix) (with 10x Hot-Taq Buffer) (with 10x Hot-Taq Buffer, 5x TuneUp solution, dntp Mix) (with 10x Hot-Taq Buffer) (with 10x Hot-Taq Buffer, 5x TuneUp solution, dntp Mix) 2, 2,

4 HelixAmp TM Hot-Taq Polymerase (Ver. 2.0) CERTIFICATE OF ANALYSIS (1603-V01R03) [MgCl 2 free] Contents HelixAmp TM Hot-Taq Polymerase (Ver. 2.0) [MgCl 2 free] Cat. No. HTBF250/HTBF250N HTBF500/HTBF500N HTBF2500/HTBF2500N Hot-Taq (2.5 units/μl) 0.1 ml 0.2 ml 0.2 ml x 5ea 10x Mg free Buffer [Hot-Taq] 1 ml 1 ml x 2ea 1 ml x 10ea 25 mm MgCl 2 1 ml 1 ml x 2ea 1 ml x 10ea dntp Mix (each 10 mm) None / 0.2 ml None / 0.4 ml None / 0.4 ml x 5ea 5x TuneUp TM solution None / 0.5 ml None / 0.5 ml x 2ea None / 0.5 ml x 10ea Blue Box 1 ea Certificate Analysis 1 ea 1 ea 1 ea Description HelixAmp Hot-Taq DNA polymerase (Ver. 2.0) is a chemically-modified form of purified Taq DNA polymerase and quite suitable for high-specific hot-start PCR, real-time PCR and multiplex PCR. The attached heat-labile chemical moiety makes Taq DNA polymerase inactive and suppresses the polymerization from non-specifically bound primers which occurs during the setting of PCR mix and first ramp-up of thermal cycling. During the first denaturation step of PCR the chemical moieties are released from Taq polymerase and the enzyme turns to be active. With the high specificity HelixAmp Hot-Taq polymerase (Ver 2.0) shows high performance in the reactions of genotyping (microsatellite or SNP), mutiplex PCR, and realtime PCR. For the maximum performance extremely pure dntps and TuneUp solutions are also included. TuneUp solution helps the DNA polymerase to efficiently amplify the problematic target region of high G+C content or structural problem. Application Hot-Start PCR Real-Time PCR Genotyping Multiplex PCR Storage buffer 50% Glycerol, 20 mm Tris-HCl (ph 8.0), 100 mm KCl, 0.1 mm EDTA, 1 mm DTT, 0.5% Tween 20, 0.5% Nonidet P-40, Concentration : 2.5 units/μl Store : -20

5 Hot-Taq Polymerase (Ver. 2.0) [MgCl 2 free] Quality Control Assay Contamination Assay for endo- and exodeoxyribonuclease `HelixAmp Hot-Taq polymerase (Ver. 2.0) is a chemical-modified form of HelixAmp Taq polymerase. HelixAmp Taq polymerase was passed from quality control assay for contamination of endo- or exodeoxyribonuclease and the bacterial host DNA host using genomic sequence-specific primer set from host bacteria. Quality authorized by Youn Taek Go Protocol Although precipitates could be arised in the 10x Buffer, they will not affect the enzyme activities 1. Recommended amount of template DNA. Human genomic DNA : 10 ~ 100 ng Bacterial genomic DNA : 5 ~ 50 ng Purified plasmid or phage DNA : 1 ~ 5 ng 2. Mix following components in a PCR tube. Template Components 10x Mg-free buffer [Hot-Taq] Volumes (μl) X μl 5 μl 25 mm MgCl 2 2 ~ 7 μl (a) dntp Mix (each 10 mm) Forward Primer (10 pmoles/μl) Reverse Primer (10 pmoles/μl) 5x TuneUp solution (b) Distilled water 1 μl 0 ~ 20 μl 1.25 units to 50 μl (a) The optimal Mg 2+ concentration should be determined empirically, but in most cases a concentration of 1.5 mm will produce satisfactory results. (b) TuneUp Solution is an additive altering the binding behavior of primer and template and can help the amplification that do not work well under standard PCR condition. Especially, TuneUp Solution can be used for the amplification of problematic template, such as high G+C content and repeat sequence regions. TuneUp Solution uses as adding into PCR reaction mixture from 0.5x to 2x.

6 Hot-Taq Polymerase (Ver. 2.0) [MgCl 2 free] 3. PCR condition Temperature & time Cycles 95, 15 min x 1 95, 20 sec Annealing Temp., 40 sec x 25 ~ 40 72, 1 min/kb (Expected size of product) 72, 5 min x 1 Annealing Temp. = T m (6 ~ 8 ) T m (Melting Temp.) = [4 x (G + C)] + [2 x (A + T)] Products Cat. No. Products Size HTBF250 HTBF250N HTBF500 HTBF500N HTBF2500 HTBF2500N [MgCl 2 free] (with 10x Mg-free Buffer, 25 mm MgCl 2 ) [MgCl 2 free] (with 10x Mg-free Buffer, 5x TuneUp solution, dntp Mix, 25 mm MgCl 2 ) [MgCl 2 free] (with 10x Mg-free Buffer, 25 mm MgCl 2 ) [MgCl 2 free] (with 10x Mg-free Buffer, 5x TuneUp solution, dntp Mix, 25 mm MgCl 2 ) [MgCl 2 free] (with 10x Mg-free Buffer, 25 mm MgCl 2) [MgCl 2 free] (with 10x Mg-free Buffer, 5x TuneUp solution, dntp Mix, 25 mm MgCl 2) 2, 2,