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Shanghai Qifa Experimental Reagent Co., Ltd., No. 6619 Chuansha Road, Chuansha Town, Pudong, Shanghai
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Shanghai Qifa Experimental Reagent Co., Ltd
KlenTaq-S sequencing enzyme
NegotiableUpdate on 05/06
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Overview
KlenTaq-S sequencing enzyme is a modified version of Taq DNA polymerase, which is a large fragment of Taq DNA polymerase and has undergone gene mutation. This enzyme has 5 '→ 3' polymerase activity, but no 5 '→ 3' exonuclease activity or 3 '→ 5' exonuclease activity.
Product Details
this company KlenTaq-SSequencing enzymes are recombinant proteins that are expressed and purified through multiple steps.
Definition of ActivityActive units refer to 72℃Below, 1×AM PCR Buffer1，30 minInternal will 10 nmole dNTPThe amount of enzyme required to add acid insoluble substances.characteristic
1. Especially suitable ddNTP Addition reaction of modified nucleotides
2. Compared to the overall length Taq DNA Polymerase, the specific activity of this enzyme has decreased
Scope of application
ddNTP Termination LawDNA Sequencing（First generation sequencing）
Pyrophosphate hydrolysis reaction extension method SNP analysis
quality control
After strict quality control testing, the product is ensured to have the highest activity and purity.
Enzyme storage buffer
20 mM Tris-HCl (pH 8.0), 100 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.5% Nonidet P40, 0.5%Tween20, and 50% glycerol.
Application Examples
The following reaction examples are 50 μlstandard PCRSystem, for reference only. actual PCRThe conditions should be optimized based on the template, primers, and target fragment size to determine the optimal reaction conditions.
1）Prepare the reaction system according to the table below
component |
volume/μl |
final concentration |
10×AM PCR Buffer 1 |
5 |
1× |
dNTPs（2.0 mM） |
5 |
0.2 mM |
primer F（10 μM） |
1.5 |
0.3 μM |
primerR（10 μM） |
1.5 |
0.3 μM |
Template |
Varable* |
/ |
KlenTaq-SSequencing enzyme（5U/μl） |
0.5 |
2.5U |
ddH2O |
Varable |
/ |
total volume |
50 |
/ |
*Template DNA dosage parameters (50 μ l reaction system):
Human genomeDNA100-1000 ng ; Microbial genomic DNA 10-100ng;PCR 产aracteristically annoyed kanyam; 1 ng- 10% of； Plasmid DNA 5-30 ng ; cDNA from RT reaction 1-5 μl 。
2）Perform the reaction under the following conditions,
95℃ |
2-5 min |
|
95℃ |
15 sec |
25-35 Cycles |
55~72℃ |
20 sec |
|
72℃ |
1kb/min |
|
72℃ |
5 min |
3）Agarose gel electrophoresis detectionPCR Product, or carry out subsequent operations according to the experimental purpose.
Precautions1.PCR Conditions and wild type Taq DNA Similar to polymerase, for specific DNA The sequencing reaction may need to be optimized.2. The amplification ability of this enzyme is higher than that of the wild type Taq DNA The polymerase has decreased,PCR The yield has decreased, and it is not recommended to use it for routine purposes PCR.3. Not available for use Taqman probe method q-PCR.
