Team:Hawaii/Notebook/2008-10- 2

From 2008.igem.org

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(New page: {{Team:Hawaii/Header}} = Things we did today = == Wetlab work == ===Verification of transformants=== :<strong> Grace</strong> [[Image:100208colonyPCR.png|right|thumb|300px|EtBr stained 2%...)
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:* Colony PCR of transformants
+
:* Colony PCR of transformants\
 +
::* Most lanes blank; lanes with bands are mostly faint. Didn't load enough PCR product? Will reload gel tomorrow with remainder of PCR product.
:* Restreaked:
:* Restreaked:
 +
::* rbs+GFP+tt #9, 10, 16, 17
 +
::* slr1+GFPf+tt #13
 +
::* nir+rbs+slr1+GFPf #1, 2, 15, 17
 +
===Plasmid Prep===
 +
:<strong> Margaret</strong>
 +
:*oriV 3, 4, & 6
 +
===Restriction Digest===
 +
:<strong> Margaret </strong>
 +
:*oriV3 E,S
 +
:*oriV4 E,S
 +
:*oriV6 E,S
 +
:*oriV11 E,S
 +
:*plac/rbs/rep 8 X,P
 +
:*pSB1A3 E,P
 +
:*oriT E,S, and another sample of oriT with X,P
 +
 +
===Culture===
 +
:<strong> Margaret </strong>
 +
:*ap1, and ap2. Because I need to digest them with E,S so that I can use as a front insert
 +
 +
===PCR Verification===
 +
:<strong> Margaret </strong>
 +
 +
:*10 more colonies of plac/rbs/rep
 +
:*check if SAP and SAP buffer is contaminated
 +
 +
 +
===Sequencing===
 +
:<strong> Margaret </strong>
 +
 +
:*oriV 3, 4, 6
 +
:*plac/rbs/rep 8 with 11 primers to cover whole sequence

Latest revision as of 04:33, 3 October 2008

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

Things we did today

Wetlab work

Verification of transformants

Grace
File:100208colonyPCR.png
EtBr stained 2% agarose gel ran at 95V for 1 hour. Two microliters of PCR product were loaded into each well.
Construct Colonies
pSB1A3, SAP'd 0
pSB1A3 to self 40
plac + rbs+GFP 42
rbs + slr1+GFP 89
rbs+GFP + tt 227
slr1+GFPf + tt 178
nir + rbs+GFP 112
nir+rbs + slr1+GFP 215
nir + rbs+GFP+tt #1 300+
nir + rbs+GFP+tt #2 103
plac + rbs+GFP+tt #1 63
plac + rbs+GFP+tt #2 11
  • Colony PCR of transformants\
  • Most lanes blank; lanes with bands are mostly faint. Didn't load enough PCR product? Will reload gel tomorrow with remainder of PCR product.
  • Restreaked:
  • rbs+GFP+tt #9, 10, 16, 17
  • slr1+GFPf+tt #13
  • nir+rbs+slr1+GFPf #1, 2, 15, 17

Plasmid Prep

Margaret
  • oriV 3, 4, & 6

Restriction Digest

Margaret
  • oriV3 E,S
  • oriV4 E,S
  • oriV6 E,S
  • oriV11 E,S
  • plac/rbs/rep 8 X,P
  • pSB1A3 E,P
  • oriT E,S, and another sample of oriT with X,P

Culture

Margaret
  • ap1, and ap2. Because I need to digest them with E,S so that I can use as a front insert

PCR Verification

Margaret
  • 10 more colonies of plac/rbs/rep
  • check if SAP and SAP buffer is contaminated


Sequencing

Margaret
  • oriV 3, 4, 6
  • plac/rbs/rep 8 with 11 primers to cover whole sequence


Discussion

Quote of the Day

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


[http://manoa.hawaii.edu/ Sponsor_UHM.gif][http://manoa.hawaii.edu/ovcrge/ Sponsor_OVCRGE.gif][http://www.ctahr.hawaii.edu Sponsor_CTAHR.gif]