Conditions and Factors Affecting PCR Pogoji in dejavniki, ki vplivajo na PCR
Factors Affecting PCR: Dejavniki, ki vplivajo na PCR:
PCR Reaction Volume Reakcijske zmesi PCR obsega
If you are increasing the PCR reaction volume proportionally to a PCR reaction that is optimized or works well, in general you will increase the amount of product with increasing volume. Če se poveča volumen reakcijske zmesi PCR v sorazmernem deležu do reakcije PCR, ki je optimizirana ali deluje dobro, v splošnem si bo povečala količina izdelka s povečanjem volumna. Exceptions to this include older mineral oil PCR machines and PCR thermocyclers without a heated-lid. Izjeme od tega vključuje starejše mineralna olja PCR, PCR, stroji in thermocyclers brez-pokrovom ogreto. In these cases, increasing the PCR reaction volume will negatively affect the PCR product and efficiency of the reaction. V teh primerih, s čimer se poveča reakcijske zmesi PCR volumna bo negativno vplivalo na produkta PCR in učinkovitosti reakcijo.
PCR Primer Design PCR Primer Design
Primer are usually chosen to be between 18-25 nucleotides in length. Primer so ponavadi izbrane, da se med 18-25 nukleotidov v dolžino. Longer primers also work 30-40 nucleotides or bp during pcr Več oligonukleotidov tudi delo 30-40 nukleotidov v PCR ali bp
A GC clamp at the 5' ends of the primers helps. A GC sponko na 5 'koncih je oligonukleotidov pomaga.
One can thus design primers with 1 or 2 nucleotides GC at the start and ends of the primers if possible Eden lahko tako načrtovanje oligonukleotidov z 1 ali 2 nukleotidov GC na začetku in konča na oligonukleotidov, če je mogoče,