Now four or more single nucleotide repeats, such as (... AAAAAA ...) 5, GC clamp: In the primer 3

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Tao Inscription: I can not say that PCR primer design expert, but reading and during graduate work has been engaged in bio-industry, karyotype for PCR primer design know little about, see the network in a number karyotype of articles on the primer design strategy is mostly copied from, There are too fraught, the special karyotype feelings and experiences on primer design are listed below, in order informative, if wrong, please point out!
Now four or more single nucleotide repeats, such as (... AAAAAA ...) 5, GC clamp: In the primer 3 'end containing a G or C, may improve the amplification effect. 6, non-target homology: The selected primers BLAST comparison, to see whether the homology. ---www.ncbi.nlm.nih.gov/BLAST/ 7, codon degeneracy: If amplify the coding region, the primer karyotype 3 'end do not terminate in the first three codons, because the first three codons easy degeneracy occurs, it will affect the amplification specificity and efficiency. 8, the annealing temperature: Tm = 4 (G + C) +2 (A + T), the estimated karyotype Tm value of the primer, the effective Tm of the primers 55 ~ 80 , its value is preferably close to 72 Tm so renaturation the best conditions. . . . .
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