Bead Cleans, Digestions, Quantifications

Dates Performed: August 30th, September 14th

PLATE A: Bead Clean Plate A 8/30/2021

• 1X bead clean (leave beads in!)

• volume=50ul

  Thawed plate A, vortexed and spun Made new 80% ethanol Followed 1X bead clean protocol (Step 5:https://docs.google.com/document/d/1iWBGgBZuXlVuiEV-A3ecxeyn9ZaY389tAqGZWksjs5Y/edit)

  Digestion Plate A Step 6 of Protocol:

  • Thawed buffer, enzymes (PST1 and EcoR1)
  • Vortexed and centrifuged
  • Made master mix of buffer and enzymes Plate A has 37 samples – n= 42 for error Plate C master mix of enzymes o 8μl Cutsmart Buffer (10X) * n = 336 o 1μl Pst1 * n = 42 o 1μl EcoR1 * n = 42 AGAIN I did not make enough and had to go back and make more master mix. Not sure what part I messed up.

Strip tube setup for plate A for master mix

Ran in Centrifuge 4 on 12 hour Digest with 2 holds

1.5X Post Digest Bead Clean Plate A 8/31/2021 Step 7 of protocol
Used PEG NaCl from 6/21/2021
Placed plate back on orbital mixer after adding 33ul of Tris HCL for minutes then placed on magnet to quantify.

Qubit Post Bead Clean

Ran 3 samples from ethanol waste and 1 from new ethanol on Qubit to see if DNA was present in either

Tapestation Post Digest and 1.5X Bead Clean (Genomic Tape)