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User:University of Washington/8 July 2008
From 2008.igem.org
BioBrick Promoter Construct Sequencing
- Because the previous forward sequencing reaction and analysis yielded high-noise results, the forward sequencing protocol was carried out a second time.
- VF2 primer (100pmol/uL) was diluted 1 to 100 with distilled water.
- 8 uL of diluted VF2 primer (1pmol/uL) was added to four Eppendorf tubes.
- 4 uL of miniprepped plasmid DNA for parts I20260, I20268, I20269, and I20270 were added to tubes of primer, after appropriate labeling of the tubes.
- Plasmid template and VF2 primer solutions were resubmitted to the UW DNA Sequencing Facility for reaction and analysis.
- The index and middle fingers were crossed.
RP4 Conjugation
·Testing DH5α and S17-1+RP4 resistance to Chloremphenicol and Ampicillin.
·Testing S17-1 to DH5α conjugation using RP4 plasmid.
·Make glycerol stock of S17-1+RP4 UW Glycerol Stocks
Non-RP4 Conjugation
·Plate out CV13(Yep13) and pDPT51
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