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Bead Clean and Quantify 59 Sea Star Libraries

Bead Clean and Quantify all Final Sea Star Libraries From X, X and X

1X Cleanup on 59 Samples

  • Placed all samples into one plate for easy cleanup
  • Layout:
  • Used multichannel for all pipetting
  • Added 15μl (should be 1X) of KAPA pure beads to each sample and pipetted up and down at least 10 times to mix beads careful to avoid bubbles
  • Placed tubes on shaker at room temp for 15 minutes
  • Placed tubes on magnet plate and removed supernatant from tubes when it was fully clear not disturbing the beads
  • Added 200μl of 80% EtOH to each tube while still the magnet not disturbing the beads
  • Removed supernatant from each tube on the magnet plate without disturbing the beads
  • Added 200μl of 80% EtOH to each tube while still the magnet not disturbing the beads
  • Removed ALL the supernatant from each tube on the magnet plate without disturbing the beads. Extra EtOH blobs were removed with p20 pipette tips
  • Resuspended beads in 16μl 10mM Tris HCl pH 8 and incubated tubes on shaker for 5 minutes
  • Placed tubes back onto the magnet stand and removed 16ul supernatant when clear to new labeled PCR strip tubes

QC

Sample Library Number Library concentration ng/ul
CHSY 003 1 52.2
CHSY 006 2 47
CPOI 005 3 40.2
CHSY 008 4 22.2
CPAB 006 5 41
CPAB2 008 6 51.4
CPBO 004 7 14
CPOI 006 8 47.6
CHSY 009 9 39
CPOI 004 10 55.4
CPAB2 001 11 25.8
CPAB2 004 12 22.8
CPBP 001 13 54
CPBO 008 14 5.4
CPBP-006 15 57.8
CPAB2 005 16 50.4
CPDB 001 17 58
CPOI 002 18 48.8
CHSB 009 19 36
CHSB 003 20 54.8
CPAB2 002 21 40.6
CPAB-001 22 60.4
Tank Escapee 23 58.2
CPAB 004 24 38.2
CPAB2 007 49 37
CPAB-003 50 50.2
CHSB 011 51 40.2
CPBO-003 52 52.8
CPDB 007 53 37.2
Chip 54 49.6
CPAB 009 55 34.6
Bash 56 52.4
CPDB 002 57 45.6
CPOI 010 58 35.4
CPDB 010 59 36.6
CPAB 002 60 32.6
CHSY 010 61 32
CPBO 006 62 37.8
CPBO 005 63 40.4
CHSY 002 64 48.8
CPOI 009 65 25.4
CHSB 004 66 28
CPBP 010 67 46.8
CPAB2 009 68 48.4
CPAB 005 69 46.6
CPDB 005 70 34.2
CPOI 008 71 47.6
CPAB2 006 72 35.4
CPOI-003 73 26.2
CPBP 004 74 32.4
CPAB 008 75 30.0
CPBP 002 76 34.8
CPBP 007 77 31
CPDP-003 78 33.8
CPAB 011 79 31
CPOI 007 80 40.4
CPAB2 003 81 30.2
CPDB 009 29 43.2
CPBP 009 43 5.4

Looks like library 14 needs to be re-done

Written on July 29, 2021