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This protocol has been adapted by the San Diego Police Department (SDPD) Crime Laboratory from the standard QIAGEN pretreatment for casework samples, and is intended for the lysis and extraction of DNA from forensic samples of fired (casings) and unfired (cartridges) ammunition.
This procedure has been used successfully for isolation of 150-250 kb BAC DNA from a mouse-BAC library cloned in pBeloBAC11 from Escherichia coli strain HB101/r. The yield of BAC DNA from 100 ml culture was typically 20-40 μg.
As starting material, 5 g soil was mixed with different amounts of Bacillus subtilis cells. Sensitivity was 5 x 103 cells/5g soil.
The procedure has been used successfully for isolation of 110 kb P1 DNA (pAdsacBII with an 80 kb insert) from Escherichia coli strain NS3529. Yield of P1 DNA was typically 10-50 µg from 500 ml culture.
The protocol describes the preservation and concentration of stool samples (in preparation for microscopic examination for intestinal parasites), using two systems from Meridian Biosciences, followed by isolation of DNA from the stool samples for pathogen detection using the QIAamp DNA Stool Mini Kit.
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