Week 3:
06/20/2011
1. Lab meeting
2. Ligate the blunted ended DNA
06/21/2011
1. Column purify the DNA
2. Make buffers for competent cells
Buffer: 10mM KOAc+80mM CaCl2-2H2O+70mM MnCl2-4H2O+10mM MgCl2-6H2O
3. Transformation again for:
Plasmids:OLE1, MSN2, TSP1, ELO1, CAS0, pSB1AK8
Vectors: prs 423-426
06/22/2011
1. Inoculation for :
pRS vectors 423-426 and pSB1AK8
2. PCR ligation products:
OLE1, MSN2, TSP1, ELO1
06/23/2011
1. Transfer E.Coli from agar stabs from iGEM delivery
BBa-K191005
BBa-K191004
BBa-K426020
BBa-I15010
Notes:
+ The agar should already have a hole from when it was stabbed. With an inoculating loop dipped into the stab, you can plate directly
onto a petri dish with the appropriate antibiotic;
+ Incubate the dish overnight at 37 C (14-16hrs)
+Pick a single colony to start up a culture
2. Gel running to test OLE1, MSN2, TSP1, ELO1, CAS0,
Observation: OLE was the only one with a clear band
06/24/2011
1. Observations of the E.Coli transformation from yesterday:
BBa-K191005 and BBa-K191004 are purple
BBa-I15010 is good
06/25/2011
1. Transform for 4 ligated genes: OLE1, MSN2, TSP1, ELO1
Notes: use 5 µL of samples
2. Inoculate for:
BBa-K191005
BBa-K191004
BBa-K426020
BBa-I15010
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