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Genomic DNA Extraction from Buffy Coat Using the Perfect gDNA Blood ,,, Mini Kit

12,00016,000 x g. Carefully remove the spin column without splashing Wash Buffer onto the bottom of the column. Place the spin column into a fresh microcentrifuge tube.
  • Observe the filter membrane to make sure it is dry before proceeding. If the filter looks shiny, spin an additional 1 to 2 minutes before placing the spin column into the fresh microcentrifuge tube.
  • Add 200 l Elution Buffer to the center of the spin column, making sure that the buffer comes into contact with the spin column filter. Incubate the sample at 70C for 3 minutes. Centrifuge for 1 minute at 12,00016,000 x g to elute gDNA. Store purified gDNA at 4C.
    • 1.5 ml of blood:
    The following protocol is intended for the researcher who wants to use only an aliquot of a whole blood sample for isolation of gDNA. An aliquot of the whole blood sample may be removed from the vial using a syringe and placed in a microcentrifuge tube. The blood is usually drawn under vacuum conditions into the vial. It is important that the aliquot of blood removed from the vial is used right away to avoid changing the properties of the blood.
    1. Transfer 1.5 ml of whole blood to a microcentrifuge tube.
    2. Centrifuge tube at 3,000 x g for 10 minutes per microcentrifuge.
    3. Continue with step 3 from the protocol outlined above.
    Gel Electrophoresis:
    200 ng (based on OD reading) was run on a 0.6% agarose gel in 1x TBE. The gel was stained in ethidium bromide. Consistency of bands as well as band size from sample to sample was evaluated.

    PCR:
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    Source:


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