Preparation of DNA Ladder Based on Multiplex PCR Technique

Figure 1

Illustrations of the PCR products for adjusting reaction system and temperature profile. 100–1000 bp fragments were amplified by multiplex PCR using 10 primer pairs in a PCR tube which have the same sense primer. (a) the annealing temperature profile from 58^∘C to 48^∘C for 30 cycles; (b) 100–500 bp fragments were amplified under the annealing temperature profile from 58^∘C to 46^∘C for 35 cycles. (c) Adjusting the amount of the ten anti-sense primers and the annealing temperature profile from 56^∘C to 46^∘C, most of target fragments were detected; (d) Raising the amount of 600–700 bp anti-sense primers and using the annealing temperature profile from 56^∘C to 44^∘C, all the target fragments were amplified and nonspecific fragments disappeared.

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