User:Karmella Haynes/Notebook/BioBrick cloning/2015/08/26

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08/26/15

  • Minipreps - CRISPR library
  • Cultures - CRISPR library (round 2)



Minipreps - CRISPR library


  • Minipreps
    • Sigma GenElute kit, elute w/ 75 μL elution sln.
  1. L34-032
  2. L34-040
  3. L34-094
  4. L34-096


  • Measure conc.'s
Sample OD260 260/280 ng/μL
1. L34-032 0.167 1.863 167.3
2. L34-040 0.026 1.908 26.4
3. L34-094 0.032 1.901 32.4
4. L34-096 0.142 1.859 142.4
  • Sequencing: Order 10437
  1. L34-032, P163
  2. L34-040 "
  3. L34-094 "
  4. L34-096 "



Cultures - CRISPR library (round 2)

  • Re-grow cultures for clones with failed sequencing runs
  • From DNASU Order 10333
    • Note: DNASU sampling from the 96-well block is top to bottom, then left to right
    • A1 = G34-001, A2 = G34-002, A3 = G34-003, etc.
  • Low phred scores (<700) generally corresponded with the lowqual reads observed in Benchling
  • Scrape bacteria from the original spot agar plates; grow as 5 mL culture for miniprep (overnight)

Clone, well (agar spot position), phred score

  1. G34-024, H3, 746
  2. G34-067, C9, 444
  3. G34-079, G10, 888


RESULTS(08/27/15)

  • All three grew



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