Week 13:
09/13/2011
1. Check quality of gDNA
2. Make HT115 + BL21 (tetO, CRISPR-GFP #7) competent cells, freeze stock
3. Miniprep PCDF+casO
C. Plate 4 Well 16O (LovTAP Composite)
4. Digest tetO/tetR
09/15/2011
1. Run a gel of all minipreped PCDF+casO
2. PCR HT115, DH5α2, DH5α3 with primers
09/18/2011
1. Inoculate the following:
BL21 tetO::GFP
BL21 tetO::GFP, CRISPR::GFP#7
BL21 tetO::GFP, CRISPR::GFP#7, cas3
HT115 tetO::GFP
HT115 tetO::GFP, CRISPR::GFP#7
HT115 tetO::GFP, CRISPR::GFP#7, cas3
HT115 CRISPR::GFP #7
09/19/2011
1. Growth test, grow all the bacteria overnight
2. PCR cas3+casO from gDNA
09/20/2011
1. Make biobrick of cas3 and casO
2. Run a gel for cas3 and casO
3. Purify cas3 and casO
4. New experiment: dilution the bacteria to 1000 and 5000 cells and plate them
09/21/2011
1. Do growth test for the following:
BL21 tetO::GFP-IPTG
BL21 tetO::GFP+IPTG
BL21 tetO::GFP, CRISPR::GFP#7-IPTG
BL21 tetO::GFP, CRISPR::GFP#7+IPTG
BL21 tetO::GFP, CRISPR::GFP#7, cas3-IPTG
BL21 tetO::GFP, CRISPR::GFP#7, cas3+IPTG
BL21 tetO::GFP, CRISPR::GFP#7, cas3+IPTG an hour later
HT115 tetO::GFP-IPTG
HT115 tetO::GFP+IPTG
HT115 tetO::GFP+IPTG an hour later
HT115 tetO::GFP, CRISPR::GFP#7-IPTG
HT115 tetO::GFP, CRISPR::GFP#7+IPTG
HT115 tetO::GFP, CRISPR::GFP#7+IPTG an hour later
HT115 tetO::GFP, CRISPR::GFP#7, cas3-IPTG
HT115 tetO::GFP, CRISPR::GFP#7, cas3+IPTG
HT115 tetO::GFP, CRISPR::GFP#7, cas3+IPTG an hour later
HT115 CRISPR::GFP #7-IPTG
HT115 CRISPR::GFP #7+IPTG
HT115 CRISPR::GFP #7+IPTG an hour later
BL21 tetO::GFP, CRISPR::GFP#7, cas3
HT115 tetO::GFP
HT115 tetO::GFP, CRISPR::GFP#7
HT115 tetO::GFP, CRISPR::GFP#7, cas3
HT115 CRISPR::GFP #7
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