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Analysis of population-level variation by sequencing PCR- amplified DNA

, : Analysis of population-level variation by sequencing PCR- amplified DNA. Molecular genetic analysis of populations: a practical approach: 159-187

The methodology for DNA amplification by PCR (polymerase chain reaction) and for sequencing PCR-amplified DNA is examined under the following headings: PCR machines (heating and cooling; controlling the cycle; software; effect of profile on yield and specificity of amplified products; and Taq polymerase); sample preparation (DNA extraction); designing primers; reaction conditions (additives that improve yield and specificity; contamination; and temperature cycle); cloning double-stranded PCR product; and preparing DNA template and direct sequencing (sample preparation; asymmetric PCR; biotinylated primers and magnetic beads; and sequencing double-stranded PCR product). In all, 11 protocols are presented for the various procedures covered.


Other references

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