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This kit contains a complete set of crude DNA extraction and PCR amplification systems, suitable for rapid genotyping of mice (Rapid Genotyping). This kit can be used to quickly release genomic DNA from mouse tails, ears, and toes. PCR amplification can be performed directly, without homogenization, disruption, overnight digestion, phenol-chloroform extraction, DNA precipitation or column purification, etc., which greatly reduces the time-consuming experiment. When using, soak the tissue in pre-added Proteinase K In the lysate, incubate at 55°C for 20 minutes and heat at 95°C for 5 minutes to inactivate Proteinase K. The lysate can be directly used as a PCR amplification template after centrifugation, and it is widely applicable to the amplification of target fragments within 2 kb after repeated testing. Suitable for multiple PCR reactions within 4 pairs of primers.
The kit is equipped with 2 × Taq Plus Master Mix (Dye Plus), including high-performance Taq Plus DNA Polymerase, dNTP and optimized buffer system. The PCR reaction only needs to add primers and templates to perform amplification, which reduces operations such as tube opening/pipetting, significantly reduces sample cross-contamination and improves detection throughput and results reproducibility. The unique protective agent allows Taq Plus Master Mix to maintain stable activity after repeated freezing and thawing. The system is premixed with electrophoresis buffer and dyes, and electrophoresis can be carried out directly after the reaction, which is convenient and quick to use. The PCR product has A at the 3'end, which can be cloned into the T vector and is suitable for ClonExpress rapid cloning kit (Vazyme #C112/C113/C114).
Special lysate based on proteinase K;
The lysate is directly used for PCR amplification without the need to extract the genome;
It is widely suitable for the amplification of target fragments within 2 kb, and is suitable for multiple PCR reactions within 4 pairs of primers.
2 × Plant Direct Master Mix should be stored at -20℃ and avoid repeated freezing and thawing; Plant Direct Lysis Buffer can be stored at -20℃ or 2℃ - 8℃ for up to 18 months.