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  • Tail Genomic DNA

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    Tatiana H. Gomez
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    This document provides instructions for extracting genomic DNA from a mouse tail sample. The process involves incubating a tail sample in an extraction buffer containing Tris, EDTA, NaCl, SDS, and proteinase K at 55C for at least 4 hours. Phenol-chloroform extraction is then used to separate and purify the DNA, followed by ethanol precipitation to concentrate the DNA. The purified genomic DNA pellet is dissolved in water and stored.

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    This document provides instructions for extracting genomic DNA from a mouse tail sample. The process involves incubating a tail sample in an extraction buffer containing Tris, EDTA, NaCl, SDS, and proteinase K at 55C for at least 4 hours. Phenol-chloroform extraction is then used to separate and purify the DNA, followed by ethanol precipitation to concentrate the DNA. The purified genomic DNA pellet is dissolved in water and stored.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as DOCX, PDF, TXT or read online from Scribd
    Flag for inappropriate content
    Download as docx, pdf, or txt
    14 views1 page

    Tail Genomic DNA

    Uploaded by

    Tatiana H. Gomez
    This document provides instructions for extracting genomic DNA from a mouse tail sample. The process involves incubating a tail sample in an extraction buffer containing Tris, EDTA, NaCl, SDS, and proteinase K at 55C for at least 4 hours. Phenol-chloroform extraction is then used to separate and purify the DNA, followed by ethanol precipitation to concentrate the DNA. The purified genomic DNA pellet is dissolved in water and stored.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as DOCX, PDF, TXT or read online from Scribd
    Flag for inappropriate content
    Download as docx, pdf, or txt
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    Tail Genomic DNA

    1. Place 0.5ml of extraction buffer with tail in each MCtube Extraction buffer (5ml):

    10mM Tris ph 8.0: 50µl of 1Mtris


    2. Incubate at 55C at least 4h. Vortex every hour.
    10mM EDTA: 100µl 0.5M EDTA
    3. Add 0.2 ml sterile water
    0,2M NaCl: 200µl of 5M NCl

    4. Add 0.3 (300µl) phenol/300µl chloroform 0,5% SDS: 125 µl of 20% SDS

    0,2 µg/ml PK: 100µl PK (10mg/ml)


    5. Vortex 30secs, centrifuge 10sec at full speed.

    6. Take the upper layer carefully, leave a thin water layer in the tube.
    Transfer it to a new tube

    7. Add 300µl phenol/ 300µl chloroform again.

    8. Vortex, centrifuge

    9. Obtain upper layer. Put in a new tube

    10. EtOH precip: Add 1/10 Vol (50µl) of 3M Na acetate Ph 5.2. Vortex

    11. Add 2vol (1ml) of ice cold 100%EtOH

    12. Store at -20C 20min. vortex. Place in ice 5 min

    13. Micro centrifuge for 20min at max speed at 4C. Tube tips facing up.

    14. Remove 100% EtOH, add 1ml 70% EtOH. Centrifuge 5 more min@4C

    15. Look for pellet. Remove supernatant. (Put waste in a tube in case you suck the pellet.)

    16. Dry pellet

    17. Dissolve dry pellet in 100µl dH2O

    18. Label

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