User:Karmella Haynes/Notebook/BioBrick cloning/2015/08/26: Difference between revisions

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Karmella's BioBrick Cloning</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Karmella's BioBrick Cloning</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
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# L34-094 "
# L34-094 "
# L34-096 "
# 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
# G34-024, H3, 746
# G34-067, C9, 444
# G34-079, G10, 888
RESULTS(08/27/15)
* All three grew




Latest revision as of 01:06, 27 September 2017

Karmella's BioBrick Cloning Main project page
Previous entry      Next entry

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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