赛默飞世尔Thermo Fisher Hydrometer

PCR引物设计和优化(PCR PRIMER DESIGN AND REACTION OPTIMISATION)

for calculation of the Tm isTm = 4（G + C） + 2（A + T）℃.Thus, the annealing temperature chosen for a PCR depends directly on length and composition of the primer（s）。 One should aim at using an annealing temperature （Ta） about 5℃ below the lowest Tm of ther pair

RNA Electrophoresis

bromide.Agarose gels have a smaller resolving power than polyacrylamide gels but a greater range of separation - from 200 bp to >50 kb using standard gels and electrophoresis equipment. RNAs up to 10 000 kb can be separated in agarose gels using pulsed

PCR基础知识[PCR basics]

and length must be chosen accordingly. (5) The melting and annealing temperatures of a primer are estimated as follows: if the primer is shorter than 25 nucleotides, the approximate melting temperature is calculated with the formula: Tm = 4(G + C) + 2 (A + T