FS Taq Mix Direct for Blood is a premixed, ready-to-use solution containing FS Taq DNA Polymerase, dNTPs, and all other PCR components, except DNA template and primers. FS Taq Mix Direct for Blood is specific for whole blood amplification. It contributes to fast, specific, sensitive and reproducible PCR by reducing the risk of pipetting errors, miscalculation and contamination. The FS Mix (2X) B can be used with conventional PCR machines. This direct protocol makes the process of complicated and time-consumng DNA extraction unnecessary, and also minimize the risk of cross-contamination.
FS Taq DNA Polymerase is the latest generation Taq-based DNA polymerase developed by Dongsheng Biotech. It possesses high amplification efficiency as does Taq polymerase, and fast elongation ability as does KOD polymerase, can be use in various kinds of PCR. The FS PCR Buffer is designed for FS Taq DNA polymerase, can be used in fast amplification reaction. FS Taq DNA polymerase has an elongation rate 2x higher than regular Taq DNA polymerase, and can shorten the amplification time by half. It has 5' to 3' polymerase activity but lacks 3' to 5' exonuclease activity, which results in a 3'-dA overhangs PCR product.
Contents
2X Taq Mix Direct Mix B; Nuclease-free water
Note: The product contains bromophenol blue for convenient gel loading of the PCR products. If you would like to leave it out, talk to us when you order the product.
Features
• Convenient: Direct for whole blood amplification PCR
• High yields of PCR products with minimal optimization
• Fast: saves time due to reduced number of pipetting steps
• Reproducible: lower contamination and pipetting error risk
• Higher sensitivity and fast compared to conventional Taq DNA polymerase
Applications
- Amplification for Whole Blood
-
High throughput PCR
• Long and Complex template PCR
Unit Definition
One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nM of dNTPs into an acid-insoluble form in 30 minutes at 70°C using hering sperm DNA as substrate.
Basic PCR Protocol
All solutions should be thawed on ice, gently vortexed and briefly centrifuged.
1. Add in a thin walled PCR tube on ice:
For a total 50μl reaction volume
|
Reagent/Sample |
Volume (50 µl rxn) |
Final concentration |
|
10x FS PCR Buffer |
25 µl |
1x |
|
Primer I |
Variable |
0.1-1 µM |
|
Primer II |
Variable |
0.1-1 µM |
|
Blood |
Variable |
0.2-2 µl* |
|
Water |
Variable to 50 µl |
N.A. |
*up to 5 µl can be used.
Recommendation for Blood Template in a 50μl reaction volume is 0.1-1μl.
2. Gently vortex the sample and briefly centrifuge to collect all drops to the bottom of the tube.
3. Overlay the sample with mineral oil or add an appropriate amount of wax. This step may be omitted if the thermal cycler is equipped with a heated lid.
4. Preform PCR using the following thermal cycling conditions.
|
Initial Denaturation |
94°C |
4 min |
|
25-35 Cycles |
94°C 55-68°C 72°C |
30s 30s 20 s |
|
Final Extension |
72°C |
2 min |