| Description |
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The Hotstart Taq Premix is a ready-to-use, 2X concentrated premix that contains all the reagents (except primers and template) need for running PCR. Simply add primers、template and water to successfully carry out PCR. Hotstart Taq Permix offers several advantages. Set up operation and time are significantly reduced. The chance of contaminating component stocks is eliminated. Reduction of reagent handling steps leads to better reproducibility.
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| Content |
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2X Hotstart Taq Premix containing: |
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‧Hotstart Taq DNA polymerase
‧dNTP mix including dATP、dCTP、dGTP、dTTP
‧7.5mM MgCl2
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| General reaction mixture for PCR (total 25μl) |
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- Set up each reaction as follows:
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| Component |
Volume / reaction |
Final conc. |
| Hotstart Taq Premix |
12.5 μl |
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Forward Primer
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variable |
0. 2~1.0μM |
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Reversed Primer
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variable |
0. 2~1.0μM |
| Template DNA |
＜1μg |
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| H2O |
up to 25.0 μl |
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- Mix the solution thoroughly by pipetting up and down.
- Program the thermal cycler according to the manufactures instructions.
- Place the tubes in the thermal cycler and start the reaction.
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Optimized Cycling Protocol |
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| Initial activation step |
10 min |
95℃ |
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Denaturation
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0.5 min |
94℃ |
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Annealing
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0.5 min |
50~68℃ |
| Extension |
1 min（1kb） |
72℃ |
| Number of cycles |
35~40 |
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| Final extension |
3 min |
72 ℃ |
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PCR performance test |
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High sensitivity of PCR is confirmed by using single copy plasmid as the template (amplified fragment: 450 bp). |
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