Dec. 4, 2014

PCR of 16S Region

  • Verifying species identity as some may have actually been juvenile C. gigas
  1. BC3_7 redo
  2. BC3_11 redo
  3. BC3_12
  4. BC3_13
  5. BC3_15
  6. BC3_16
  7. BC3_17
  8. BC3_18
  9. OR2_3
  10. OR2_4
  11. OR2_5
  12. OR2_6
  13. OR2_7
  14. OR2_8
  15. OR2_9
  16. OR2_10
  17. OR2_11
  18. OR2_12
  19. OR2_15
  20. OR2_16
  21. OR2_17
  22. OR2_18
  23. OR2_19
  24. OR2_20
  25. OR3_1
  26. OR3_2
  27. OR3_3
  28. OR3_5
  29. OR3_6
  30. OR3_9
  31. OR3_10
  32. OR3_12
  33. OR3_13
  34. OR3_15
  35. OR3_17
  36. OR3_18
  37. WA9_2
  38. WA9_3
  39. WA9_4
  40. WA9_5
  41. WA9_6
  42. WA9_7
  43. WA9_8
  44. WA9_9
  45. WA9_10
  46. WA9_11
Make Master Mix 1
1x          51x
Water        6 ul      306ul
dNTPs      .5ul       25.5ul
R primer    2.5 ul   127.5
F primer    2.5ul     127.5

Master Mix 2
1x            51x
Water          9.17ul       467.67
10x buffer   3.2 ul        163.2
Taq             .13             6.63

Add 1 ul DNA to eac tube
 
Run out gels
  • CA2_1,3,4,5,7 (6/2)
    • redo all
  • CA3_1,2,3,4,5,6
    • 5:3
      6:3
    • redo CA3_1,2,3,4
  • WA12D_1,2,3,4,5,6
    • 1: 0
      2: 3
      3: 4
      4: 3
      5: 5
      6: 0
    • redo WA12D_1,6
  • WA10T_1,2,4,5,6,7
    • 1: 5
      2:4.5
      4: 5
      5: 0
      6: 5
      7: 0
    • redo WA10T_4,6
  • CA6_1,3,7,8,9 redos
    • 1: 2 (low, deg)
      3: 3(low)
      7: 3(low,deg)
      8: 5
      9: 0
  • WA9_2,3,7 redos
    • 2: 2.5(low)
      3: 0
      7: 1
  • OR1_1,2 redos
    • 1: 5
      2: 3.5(low)
  • CA7_6 redos
    • 6: 3.5(low)
  • BC3_12,13,15,16,17,18
    • 12: 5
      13: 4(bright, some smear)
      15: 2(low)
      16: 5
      17: 3.5(smear)
      18: 3.5(very very bright, smear)
  • Note: Re-extract CA6_1,9, WA9_3,7, BC4
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