Monday 9/28/15

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Do AlfI digestion of ethanol precipations from last week.
  1. SS2_3
  2. HC1_1
  3. NF1_1
  4. NF1_2
  5. SS2_6
  6. SS2_7
  7. SS2_8
  8. HC1_5
  9. HC1_6
  10. HC1_7
  11. HC1_8
  12. HC1_9
  13. HC1_10
  14. NF1_4
  15. NF1_5
  16. NF1_6
  17. NF1_7
  18. NF1_8
1x
20x
10x buffer R
1.2 uL
24
150 uM SAM
.8
16
AlfI (2 U/uL)
.5
10
water
1.5
30

Did ligation of same samples. Left for 2.5 hours at 16degC and did 10 minutes at 65degC following Matz protocol.

  • 1st make new adaptors.
  • Adaptor 1
    11x
    20x
    5ILL-NR
    .6 uL
    1.1
    Anti-ILL
    .6 uL
    1.1
    NFW
    58.8 uL
    106.91
    Adaptor 2
    11x
    3ILL -NR
    .6 uL
    Anti-ILL
    .6 uL
    NFW
    58.8 uL
    Master mix
    1x
    20
    2 uM Adaptor 1
    5 uL
    100
    2 uM Adaptor 2
    5 uL
    100
    T4 ligase
    1 uL
    20
    T4 ligase buffer with 10 mM ATP
    4 uL
    80
    NFW
    25
    500

    40 uL master mix + 10 uL digestion product

1 thought on “Monday 9/28/15”

  1. Pingback: Wednesday 10/14/15 | Getting My Genes Wet

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