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HotStarTaq PCR Handbook

Amino acid Number of codons
Met, Trp
Cys, Asp, Glu, Phe, His, Lys, Asn, Gln, Tyr
Ile
Ala, Gly, Pro, Thr, Val
Leu, Arg, Ser 1
2
3
4
6
3. Number of cycles

A cycling program usually consists of between 25 and 35 cycles, depending on the number of copies of the starting template. Too many cycles do not necessarily lead to a higher yield of PCR product; instead they may increase nonspecific background and decrease the yield of specific PCR product. Table 10 provides a general guideline for choosing the number of cycles.

Table 10. General guidelines for choosing the number of PCR cycles


*Refer to Table 6 to calculate the number of molecules. When starting with cDNA templates, it is important to take into account the efficiency of reverse transcription in cDNA synthesis, which is on average 1030%.
Refers to single-copy genes.

4. Sensitive PCR assays

PCR can be performed to amplify and detect just a single copy of a nucleic acid sequence. However, amplification of such a low number of target sequences is often limited by the generation of nonspecific PCR products and primerdimers. The combin
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