Cat.No.：A02041A
Price: ￥12
Specifications：
This product is a pre-mixed PCR reaction system. The pre-mixed components include Taq DNA polymerase, Mg²⁺, dNTPs, and auxiliary components that have both stabilizing and enhancing functions. The ready-to-use design significantly simplifies the operation process, which not only shortens the reaction preparation time but also effectively reduces human operational errors and the risk of external contamination. The optimized MasterMix formula can ensure high yield of amplification products, and at the same time, it has excellent batch-to-batch reproducibility and stability.
The system does not add electrophoresis indicator dyes. Users can add an appropriate amount of loading buffer before electrophoresis according to their experimental needs. The 3' end of the amplification product has a prominent "A" tail, which can be directly used for T/A cloning. This product has stable performance and strong adaptability, and is widely applicable to conventional molecular biology experimental scenarios such as DNA fragment PCR amplification and sequence determination.
After testing, no exogenous nucleases were detected; the PCR method showed no residual host DNA; it can effectively amplify single-copy genes in various genomes.
| Cat. No. | Component | Specification | Quantity |
| B01031A | Fast Taq PCR Master Mix (2×) | 1 mL | 1 |
| B01031A | Fast Taq PCR Master Mix (2×) | 1 mL | 5 |
Stored and transported at -20℃.
This product is mainly applicable for experiments such as PCR amplification of DNA and DNA sequence determination.
Compatible template types
This product is suitable for regular DNA (genomic DNA ≤ 15 kb, cDNA ≤ 10 kb, plasmid ≤ 15 kb), high GC/AT content (GC content 24% - 79%), animal and plant lysates, and bacterial cell culture/spot. If using yeast cell culture/spot as the template for amplifying long fragments, the extension time can be prolonged (5 - 10 seconds per kb) or a preliminary rough lysis can be performed first.
Low amplification efficiency or no band formation
The suggestions are as follows for troubleshooting in sequence: ① Check if the primers have degraded and if the design is specific (BLAST can be used for verification); ② Verify if the template is fresh. Avoid repeated freezing and thawing. The crude product may contain inhibitors (properly dilute them), and the cDNA needs to ensure the RNA quality; ③ Check if the enzyme has lost its activity (replace with a new enzyme); ④ Check if the reaction system has been prepared accurately (reprepare it); ⑤ Program parameters: Verify the accuracy of the denaturation temperature, optimize the annealing temperature (gradient), and confirm that the extension time is sufficient.
Low production volume
The main reasons are primer degradation or poor template quality (it is recommended to prepare fresh); if the template is insufficient, the first amplification product can be diluted in a ratio and then amplified again. In terms of the procedure, the annealing temperature, extension time, or the number of cycles can be optimally adjusted in a gradient manner.
Nonspecific amplification
This is usually caused by non-specific binding of the primers or the formation of dimers, which can be mitigated by reducing the primer concentration or redesigning the primers. Impure templates, contamination, and excessive amounts are all important influencing factors. It is recommended to purify and adjust the dosage according to the instructions. Procedure adjustment: If the background band is smaller than the target band, increase the annealing temperature and reduce the number of cycles; if the background band is larger, shorten the extension time and reduce the number of cycles.
Diffusion or tailing of the electrophoretic bands
Ensure that the gel is completely melted; check if the primers have degraded; avoid template degradation or excessive amounts (by performing electrophoresis tests). If amplifying long cDNA fragments, ensure the quality of the reverse transcription product (random primers can be discarded for re-experiment). Additionally, optimizing the annealing temperature gradient is helpful for improvement.
Blank control shows a band
It may be due to homology between the primers and non-target sequences, or due to product contamination (when the size of the contaminant band is consistent with the target band). It is recommended to replace the Mix, water or primers, and perform the operation in a laminar flow hood to prevent aerosols and cross-contamination. If there is contamination of small fragments of nucleic acids, nested PCR can be attempted to eliminate it.
