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.