q-PCR时引物设计问题

不倒翁

是的 Rules:
# j% }9 U5 a  t% j. z! f1.        Amplicon size should be ranged from 90bp to 180bp;* |: O, ^0 k4 D, U/ Y+ k
2.        Oligo length should be ranged from 17 to 20bp, normally;6 W" f  y" N$ |( p$ |8 i
3.        Ideally, the TM value should be 60℃, but 59℃ to 61℃ is acceptable;
* E7 B& X2 ]6 x) Y4 F4.        The TM value of one pair primers should not be different at more than 1℃. }) X- p& n! p5 O* i
5.        GC content should be 45% to 55%
- M, }7 I4 o3 d6.        The oligo designed should be located in different exons./ \& s; P+ g9 F" W- M6 d