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Genomic DNA Sample Preparation and Analysis with PstI Digestion and PCR Amplification

This protocol outlines the preparation and analysis of genomic DNA using the PstI restriction enzyme. It includes steps for adaptor ligation using biotinylated adaptors for multiple samples, MIxture components, and MboI digestion. The method also details purification with magnetic beads coated with streptavidin and subsequent PCR amplification using locus-specific primers with fluorescent dyes. Finally, it covers the measurement of PCR product concentrations through fluorescent intensity analysis, allowing for precise evaluation of the DNA fragments generated.

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Genomic DNA Sample Preparation and Analysis with PstI Digestion and PCR Amplification

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  1. P P C TGCA G C TGCA G G ACGT C G ACGT C (Mix) 1 : 1 : 0.5 1) genomic DNA 2) Pst I digestion 3) Adaptor ligation (adaptor) (genomic DNA) PT1 adaptor biotinylated- adaptor sample A (reference) ACGT PT2 adaptor sample B ACGT PT3 adaptor sample C (reference) ACGT 4) Mixture each sample & Mbo I digestion M GATC GATC GATC 5) Purification with the magnetic beads coated with streptavidin & PCR amplification Dynabeads- streptavidin locus-specific primer fluorescent dye-labeled adaptor primer 6) Measurement of concentration of the PCR product PT2 PT1 PT3 fluorescent intensity length (bp)

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