Team:Hawaii/Notebook/2008-09-18

From 2008.igem.org

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(New page: {{Team:Hawaii/Header}} = Things we did today = == Wetlab work == ===Plasmid prep=== :<strong> Grace</strong> :* Inoculated TB+sp<sub>100</sub> with BB-pRL1383a for plasmid prep ===PCR t...)
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===Transformation===
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<strong>Margaret</strong>
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:*transformed ligation products in DH5-alpha
== Drylab Work ==
== Drylab Work ==

Revision as of 03:57, 20 September 2008

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Notebook (t) Meetings (t)

Things we did today

Wetlab work

Plasmid prep

Grace
  • Inoculated TB+sp100 with BB-pRL1383a for plasmid prep

PCR to verify plasmid preps

Grace
  • PCR of 3A:nrsg and 3A:nrpg13-8 plasmids

Restreaked BB-pRL1383a plate

Grace

Transformation

Margaret

  • transformed ligation products in DH5-alpha

Drylab Work

Sequencing Analysis

Grace
  • Correct:
  • C0012V (sequence to see what it was): EcoRI/XbaI/SpeI/PstI sites present. Insert aligns to sequence for lacI (C0012 insert, vector was uncut)
  • C0012V-SAP'd (sequence to see what it was): EcoRI/XbaI/SpeI/PstI sites present. Insert aligns to sequence for lacI (C0012 insert, vector was uncut)
  • slr1+GFPf
  • Incorrect:
  • 3A:nrp: EcoRI/PstI sites present. 82bp insert aligns to sequence for E. coli flagellar biosynthesis RNA polymerase sigma factor
  • 3A:nrpg12-2: EcoRI/PstI sites present. 759bp insert aligns to sequence for E. coli fumarate reductase (anaerobic) catalytic and NAD/flavoprotein subunit
  • 3A:nrpg12-6: EcoRI/PstI sites present. 899bp insert aligns to sequence for E. coli protein C5 component of RNAse P
  • 3A:nrs2: EcoRI/PstI sites present. 85bp insert aligns to sequence for E. coli diaminopimelate decarboxylase, PLP-binding
  • 3A:nrs3: SpeI/PstI sites present. Insert aligns to pilA sequence and pSB1A3 transcriptional termination stem loop
  • 3A:nrsg10: SpeI/PstI sites present. Insert aligns to pilA + GFPf
  • pilA+GFPf12: GFPf only
  • plac+rbs1 (sequence to see what it was): EcoRI/XbaI/SpeI/PstI sites present. rbs (B0034) only
  • plac+rbs2 (sequence to see what it was): EcoRI/XbaI/SpeI sites present. Insert showed 97% similarity to RP4 origin of replication
  • plac+rbs12 (sequence to see what it was): EcoRI/XbaI/SpeI/PstI sites present. Insert is GFP only.
  • Resequence:
  • plac+rbs (no read -- only 5bp sequenced ?!?)
  • pilA+GFPf13 (no read -- only 5bp sequenced ?!?)

Discussion

  • Please throw away old plates in 4C fridge. Some have fungi growing on them already.

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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