PCR teaching kit (including templates and primers)
This kit contains all the reagents required to complete the PCR reaction, and can be used for teaching and research in PCR experiments. The template is plasmid DNA; Primers are upstream and downstream primers designed based on the plasmid DNA sequence; two × Taq PCR MasterMix includes Taq DNA polymerase, dNTPs, MgCl2, reaction buffer, enhancer and optimizer for PCR reaction, and stabilizer. When in use, PCR reaction can be carried out by adding the necessary primers and templates for the experiment, which can minimize human error and has the advantages of fast, simple, and good stability. After using this reagent kit for PCR reaction, the PCR product size should be 2000bp.
Component specification concentration
Template 30 μ L 10ng/ μ L
Upstream primer (2000 FW) 30 μ L 10 μ M
Downstream primer (2000 RV) 30 μ L 10 μ M
two × Taq PCR MasterMix (with dye) 2 × five hundred μ L-
Deionized water 1mL-
Storage: -20 ℃, valid for one year.
Completely melt the template, primer, and 2 in an ice bath × Taq PCR MasterMix, mix well and quickly shake and centrifuge to collect the solution at the bottom of the tube.
Prepare the reaction system in a 0.2mL PCR tube according to the following table:
Component 50 μ L reaction system dosage
Upstream primer (2000FW) 10 μ M 1 μ L
Downstream primer (2000RV) 10 μ M 1 μ L
Template 1 μ L
two × Taq PCR MasterMix 25 μ L
Water 22 μ L
Collect the reaction solution to the bottom of the tube by rapid centrifugation.
If the PCR instrument is not heated by a hot cap, add 25 more μ L mineral oil.
Perform the following program on the PCR instrument:
Step Temperature Time Cycles
Initial denaturation at 94 ℃ for 5 minutes 1
Denaturation 94 ℃ 30sec 30
Annealing at 54 ℃ for 30 seconds
Extend 72 ℃ for 2 minutes
Final extension at 72 ℃ for 5 minutes 1
After the reaction, take 5mL of reaction solution and conduct electrophoresis on 1% agarose gel to observe the banding.
