Monday 10/26/15

DNA Extractions for Common Garden Project

I ran out of columns in the EZNA Mollusc kit and the new one I ordered hadn’t come in yet. Since I’m trying to finish all of the South Sound and Hood Canal extractions before I leave on Sunday, I decided to do one set of extractions with the Qiagen DNeasy Kit. I’ve used this kit successfully for my population structure project. Annoyingly, I grabbed the wrong set of samples and ended up re-extracting ones I had done previously. I guess the more DNA, the better. These are notated with a B in the sample sheet. Used Qubit to get DNA concentrations.

  1. SS1_1: 18.2
  2. SS2_9: 61.6
  3. SS2_10: 14.6
  4. SS2_11: 21.8
  5. SS2_12: 94
  6. SS2_13: 24.4
  7. SS2_14: 276
  8. SS2_16: 16.2
  9. SS2_17: 33.6
  10. SS2_18: 16.7
  11. SS2_19: 10.8
  12. SS3_3: 94.8
  13. SS3_4: 41.2
  14. SS3_7: 19.9
  15. SS3_8: 24.6
  16. SS3_9: 41.2
  17. SS3_10: 20.8
  18. SS3_11: 27.2
  19. SS3_13: 19.7
  20. SS3_14: 88
  21. SS3_15: 123
  22. SS3_16: 126
  23. SS4_1: 19.2
  24. SS4_4: 15.5

Since today felt like a wasted day, I decided to set up some extractions to finish tomorrow morning. I was also out of Qiagen columns, however, so looked into regenerating some of the EZNA columns. Based on references here and here I added 500 uL of 1 M HCL to used EZNA columns and left them overnight. I made sure to only reuse columns that had previously been used for South Sound extractions just in case there is any DNA carry-over.

  1. SS2_15: 23.5
  2. SS3_12: 81.2
  3. SS3_18: 7.92
  4. SS3_19: 13.9
  5. SS3_20: 48
  6. SS3_21: 19.5
  7. SS4_2: 40
  8. SS4_3: 78.8
  9. SS4_5: 24.7
  10. SS4_6: 26.8
  11. SS4_7: 50.8
  12. SS4_8: 20.4
  13. SS4_9: 11.8
  14. SS4_10: 11.3
  15. SS4_11: 18.2
  16. SS4_12: 35.4
  17. SS4_13: 31.4
  18. SS4_!4: 30.3
  19. SS4_15: 18.3
  20. SS4_16: 26.6
  21. SS4_17: 24.7
  22. SS4_18: 38.7

Left tissue in lysis buffer at 10:30pm at 56degC.

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One thought on “Monday 10/26/15

  1. Pingback: Tuesday 10/27/15 | Getting My Genes Wet

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