| gggc ggccgtttac ccatacgatg ttcctgacta tgcgggctat ccctatgacg tcccggacta tgcaggatcc tatccatatg acgttccaga ttacgctccg gccgcCCTTA ACGTGAGTTT TCGTTCCACT GAGCGTCAGA CCCC
| R1123 |
Strain XL1-blue carrying vector pHSS6. |
| R1236/B211 |
Strain RDP146 (F- recA' dlac-pro) rpsE; spectinomycin resistant) with plasmid pLB101 (pACYC184 with tnpA; active transposase, chloramphenicol resistant)(F. Heffron) |
| #111/B428 |
Strain RDP146 with pOX38 F factor derivative carrying mTn3 derivative mTn-3xHA/GFP (GFP, URA3, tet; tetracycline resistant) |
| #70/B425 |
Strain NG135 (K12 recA56 gal-delS165 strA; streptomycin resistant) with plasmid pNG54 (pACYC184 with mTn3 res and tnpR seqs; active resolvase, chloramphenicol resistant)(N. Grindley) |
| B227 |
Strain DH5-alpha carrying pB227/GAL-cre (amp, ori, CEN, LEU2) (B. Sauer) |
Vector used:
The accession for pHSS6 is M84115
Antibiotics used:
| Tetracycline HCl, Tet (Sigma T3383) |
12 mg/ ml in 50% ethanol. Use at 3 ug/ml (Tet3) |
| Kanamycin, Kan (Sigma K800) |
10 mg/ ml in water. Use at 40 ug/ml (Kan40) |
| Chloramphenicol, Cm (Sigma C0378, I think) |
34 mg/ml in ethanol. Use at 34 ug/ml (Cm34) |
| Streptomycin, Sm (Sigma S6501) |
10 mg/ml in water. Use at 50 ug/ml (Sm50) |
| Ampicillin, Amp (Sigma A9518) |
50 mg/ml in water. Use at 50 ug/ml (Amp50) |
NB. When only a few plates of each type are used, it's convenient to chop an LB plate up with a sterile toothpick, put the bits in a sterile flask, and melt the agar by microwave. Add appropriate amounts of antibiotic and repour plates.
上一篇：Methods for use with the mTn-lacZ/LEU2-mutagenized library 下一篇：Protocols for shuttle mutagenesis/epitope-tagging of a yeast gene with mTn-4xHA/
共3页: 上一页 [1] [2] 3 下一页 |
