Team:Hawaii/Notebook/2008-08-13

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(Difference between revisions)
(→Wetlab work)
(→Sequencing)
Line 43: Line 43:
:* Checked sequencing results returned from CORE Hawaii
:* Checked sequencing results returned from CORE Hawaii
-
 
+
::* Confirmed:
 +
:::* B0015
 +
:::* B0030
 +
:::* B0034 (one bp off)
 +
:::* nir
 +
:::* slr
 +
::::* slr1 = slr2  huh?
 +
::* Other:
 +
:::* nir+rbs
 +
::::* No rbs present; only nir
 +
:::::* Huh? We ligated nir INTO rbs
 +
:::* I14032+rbs
 +
::::* No plac present; only rbs
 +
:::* BB-pRL1383a
 +
::::* E0040 (GFP) went in successfully, not J33207
 +
:::::* WTF? Our plasmid preps were/are mislabeled
 +
:::::* Redo ligation to get blue/white screen?
 +
:::* GFP+tt (reverse only)
 +
::::* Low quality read; GFP is not present (segment too small)
 +
:::* GFPf+tt (reverse only)
 +
::::* lacZ fragment present; no tt
 +
:::::* Huh? We ligated the part INTO tt
 +
::::* GFPf+tt and J33207+tt samples switched?
 +
:::* J33207+tt
 +
::::* Part did not go in; tt only
= Discussion =
= Discussion =

Revision as of 19:00, 13 August 2008

Projects Events Resources
Sponsors Experiments Milestones Protocols
Notebook (t) Meetings (t)

Things we did today

Wetlab work

Verification of Transformants

EtBr stained 2% agarose gel ran at 95V for 1 hour. Five microliters of PCR reaction were loaded into each well.
Grace
Construct Colony forming units
slr1 + GFPf 69 + 1 cluster of colonies
pilA + GFPf 17
nir+rbs + GFP 0
nir+rbs + slr1 0
nir+rbs + pilA 1
plac+rbs + GFP 0
plac+rbs + slr1 0
plac+rbs + pilA 0
  • Colony PCR of transformants and restreaked colonies from yesterday
  • Ran on 2.5% agarose gel at 95V for 1 hour

Drylab Work

Sequencing

Grace
  • Checked sequencing results returned from CORE Hawaii
  • Confirmed:
  • B0015
  • B0030
  • B0034 (one bp off)
  • nir
  • slr
  • slr1 = slr2 huh?
  • Other:
  • nir+rbs
  • No rbs present; only nir
  • Huh? We ligated nir INTO rbs
  • I14032+rbs
  • No plac present; only rbs
  • BB-pRL1383a
  • E0040 (GFP) went in successfully, not J33207
  • WTF? Our plasmid preps were/are mislabeled
  • Redo ligation to get blue/white screen?
  • GFP+tt (reverse only)
  • Low quality read; GFP is not present (segment too small)
  • GFPf+tt (reverse only)
  • lacZ fragment present; no tt
  • Huh? We ligated the part INTO tt
  • GFPf+tt and J33207+tt samples switched?
  • J33207+tt
  • Part did not go in; tt only

Discussion

Quote of the Day

History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson


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