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Genomic DNA Extraction from Buccal Swabs Using the Perfect gDNA Blood ,,, Mini Kit

Laura Pollock and Jennifer Halcome
Eppendorf 5 Prime, Inc. Introduction

Buccal swabs are the least invasive way of collecting gDNA from humans, and also have the added benefit of minimizing the exposure to blood-borne pathogens. Buccal swabs are quickly becoming the method of choice when obtaining specimens for DNA testing. The swabs are easy to use and readily available. Cells collected on buccal swabs do not require special storage conditions and the DNA remains useable after years of storage. The Perfect gDNA Blood Mini Kit provides a fast and consistent method of isolating gDNA from buccal swabs that can subsequently be used in PCR1. The Perfect gDNA kit has been validated for use with several types of swabs, and will most likely work with others. The resulting gDNA is suitable for use in the main downstream applications such as PCR.

Materials
  • Perfect gDNA Blood Mini Kit
Perfect gDNA Blood Mini Kit
  • Ethanol (95100%); pipette tips; microcentrifuge; Eppendorf Thermomixer, water bath or heat block at 70C. Buccal swabs:
  • Cotton (Puritan)
  • Dacron (Fitzco)
  • C.E.P. (Life Technologies)
1x PBS (sufficient amounts are supplied for blood work, but additional amounts may be needed for buccal swab applications)

Method

Preparation of cheek cells and isolation of gDNA:

  1. Collect samples by scraping a buccal swab firmly against the ins
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Prepared by multi-temperature ethanol fractionation with extensive charcoal treatment and dialysis.• Assay: ≥ 95 % (HPCE)• pH ~5.2 (10 mg/mL in 0.15 M NaCl)...
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