2×RoomTop Fast Taq PCR MasterMix
This product is made with 2 × Based on Fast Taq PCR MasterMix and integrated with a new stabilizer, a premixed reagent has been developed with high storage stability. It can be stored at room temperature of 25 ℃ for 2 months and 4 ℃ for 6 months, and still maintains excellent amplification performance (Figure 1).
2 × RoomTop Fast Taq PCR MasterMix continues the fast and efficient advantages of the Fast series, extending 1kb at 72 ℃ for 15 seconds, significantly reducing the time required for PCR amplification and achieving the goal of shortening the entire PCR reaction time.
2 × RoomTop Fast Taq PCR MasterMix can achieve better amplification results for genomes, high GC, and complex templates. two × RoomTop Fast Taq PCR MasterMix offers two forms for you to choose from: fast loading type (with high-purity blue dye) and ordinary type without dye. Rapid sample loading can be performed directly by electrophoresis after PCR reaction, saving time. After testing, the addition of high-purity dyes did not affect the PCR reaction.
Product features:
1. PCR identification reagents that can be placed on the experimental platform.
2. Store at room temperature for 2 months and at 4 degrees Celsius for 6 months without affecting amplification performance.
3. The amplification speed of Fast Taq is more than four times that of Taq, and long fragment amplification is more time-saving.
4. It can be directly used for PCR identification experiments such as colonies and bacterial fluids.
5. The amplification effect of genomic DNA and complex templates is better.
Product use: Conventional PCR identification; Small fragment DNA cloning; PCR product plus A; Long fragment amplification
Usage suggestions:
1. The PCR product amplified using this reagent has a prominent "A" base at the 3 'end, which can be directly cloned into the T vector.
2. Please prepare the PCR reaction solution on ice and place it on the PCR instrument for testing. This cold start method can increase the specificity of PCR amplification and obtain good PCR results.
3. When using PCR to clone the target fragment, in order to reduce random mutations during the PCR process, the template size can be appropriately increased to reduce the PCR cycle (such as 20 to 25 cycles).
Application examples:
Legend 1) At 50 μ In the l system, after storing at room temperature for different times, 2 × RoomTop Fast Taq PCR Mix reagent for amplification of a 2.1kb fragment of the mouse genome
Lane M: DNA Ladder 2000 Plus Lane 1:0 weeks
Lane 2: 2 weeks Lane 3: 4 weeks
Lane 4: 6 weeks Lane 5: 8 weeks
Lane 6: 10 weeks Lane 7: 12 weeks
Common reaction systems (50 μ l) :
Component volume
two × PCR MasterMix * 25 μ L
Upstream primers 0.2-1.0 μ M (final concentration)
Downstream primers 0.2-1.0 μ M (final concentration)
Template 1-50ng (plasmid)
10ng-1 μ G (genome)
DdH2O to 50 μ L
*The final concentration of Mg2+is 2mM
Common PCR cycles:
When the amplified fragment is less than 3K:
94 ℃ for 10 minutes
30 cycles at 94 ℃ for 20 seconds
57 ℃ for 20 seconds
72 ℃ adjusted according to product length, 15 seconds/kb
72 ℃ for 3 minutes
4 ℃ insulation
When the amplified fragment is ≥ 3K (recommended primer length ≥ 30bp):
94 ℃ for 10 minutes
30 cycles at 94 ℃ for 5 seconds
68 ℃ adjusted according to product length, 15 seconds/kb
72 ℃ for 3 minutes
4 ℃ insulation
