Week 12:
08/29/2011
1. re-PCR for CASO in PCDF vector
2. Inoculate DH5α, BL21 and HT115 cells in 5mL LB
3. Purified CAS3 (#6,8,9,12)
4. Put CRISPR-tetR into PCOLA (KanR) instead of tetR (CMR)
08/30/2011
1. For CASO
gDNA isolation of DH5α and HT115 cells
PCR CASO from gDNA with primers: AP1+CASOR
2. Inoculate PCOLA+PCDF
3. For CAS3
4. Transform cas3 into tetO+CRISPR-GFP cells. Notes: use controls
5. Gel verify casO
6. Make competent cells of tetO+GFP+CRISPR+cas3
7. Inoculate 6 culture tubes by IPTG or Without IPTG
tetO+GFP+CRISPR+cas3 (#12, 9, 6)
tetO+GFP+CRISPR
8. Transformation teto-GFP or CRISPR-GFP (# 2,4,5,6,7)or tetO-GFP and CRISPR-GFP (#1,2,3,4,5,6,7) into HT115 cells
09/01/2011
1. Inoculation of HT115 with/without IPTG
2. Freeze down both tubes of tetO::GFP CRISPR-GFP (B4) and A1, B1, C1
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