Team:Hawaii/Notebook/2008-10-21

From 2008.igem.org

(Difference between revisions)
(Insertion of lac-GFP device into BB-pRL1383a)
(Wetlab work)
 
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:* Plated 50 μl cells + SOB on each LB+sp plate; plated 20 μl untransformed cells + SOB on each control plate
:* Plated 50 μl cells + SOB on each LB+sp plate; plated 20 μl untransformed cells + SOB on each control plate
:* Incubated at 37C for 2 days
:* Incubated at 37C for 2 days
 +
 +
===Construction of GFP secretion device===
 +
:<strong>Krystle</strong>
 +
:* Gel purified overnight restriction digested nir+rbs
 +
::* Loaded all of restriction digest into a 2% gel, which appeared blank after running for 1.5 hours
 +
:* Re-PCR nir+rbs and slr+gfpf from plasmid prep
 +
:* Restriction digest
 +
::*nir+rbs with EcoRI and SpeI
 +
::*slr+gfpf with XbaI and PstI
= Discussion =
= Discussion =

Latest revision as of 01:05, 30 October 2008

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Things we did today

Wetlab work

Insertion of lac-GFP device into BB-pRL1383a

EtBr stained 0.8% agarose gel ran at 60V for 90 minutes. Thirty microliters of the RE digest reaction were loaded into each well.
EtBr stained 0.8% agarose gel ran at 60V for 90 minutes. Ten microliters of PCR product were loaded.
Grace
  • Ran RE digests on gel
  • Bands were all incorrect size
  • Reinoculated TB+sp100 for plasmid prep tomorrow
  • PCR of J04430 from filter paper to verify part; used H/B primers

Team:Hawaii/Antibiotic test for BB-pRL1383a

Grace
  • Made LB+spvariable plates with 50 μl 20X X-gal
  • Transformed 100 μl DH5α with 2 μl BB-pRL1383a (4 ng)
  • Plated 50 μl cells + SOB on each LB+sp plate; plated 20 μl untransformed cells + SOB on each control plate
  • Incubated at 37C for 2 days

Construction of GFP secretion device

Krystle
  • Gel purified overnight restriction digested nir+rbs
  • Loaded all of restriction digest into a 2% gel, which appeared blank after running for 1.5 hours
  • Re-PCR nir+rbs and slr+gfpf from plasmid prep
  • Restriction digest
  • nir+rbs with EcoRI and SpeI
  • slr+gfpf with XbaI and PstI

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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