An automatic micropipette with a filter tip is an indispensable tool for PCR experiments
Oligonucleotide primers synthesized by an automatic DNA synthesizer can be used in standard PCR without further purification. If the primers are purified by commercially available resin column chromatography (such as NENSORB; NEN Life Science products) or denatured polyacrylamide gel purification, the efficiency of amplifying single-copy sequences from mammalian genome templates will be higher (in Chapter 2, Scheme 3) in).
● Template DNA. The template can be a DNA fragment, prepared genomic DNA, recombinant plasmid, or any other sample containing DNA. The template DNA was dissolved in 10mmol/L Tris-Cl (pH 7.6) containing a low concentration of EDTA (<0.1mmol/L).
● Heat-resistant DNA polymerase. Taq DNA polymerase is a standard and suitable enzyme suitable for most forms of PCR amplification stages. However, when the extension of mismatched primers at the 3'end is suspicious, the thermostable DNA polymerase with 3'→5' correcting activity (Chiang et al. 1993) can be given priority (see the section "Thermostable DNA polymerase" and information on the previous page). Column "Taq DNA polymerase").
Taq DNA polymerase is stored in a storage buffer containing 50% glycerol. The solution is quite viscous, making it difficult to pipette accurately. The best way is to use a centrifuge to centrifuge the centrifuge tube containing the enzyme at a high speed of 4'C for 10 seconds, and then use a positive displacement pipette to absorb the required amount of enzyme.
●Automatic pipette. An automatic micropipette with a filter tip is an indispensable tool for PCR experiments. The disposable filter tip with a water barrier prevents the sample liquid from accidentally flowing into the micropipette, and at the same time reduces the possibility of cross-contamination between PCR and DNA samples during the experiment.
Tips with filter elements can be purchased from several commercial companies (for example, tips with filter elements can be purchased from Yongyue Medical Technology (Kunshan) Co., Ltd.).
