Resources
All resources stored at -20°C in the freezer in 5C8, unless otherwise stated.
Vectors
Please take one 10 µg aliquot.
| pFastbacHTa | blue tubes, box 6 |
| pFastbacHTb | yellow tubes, box 6 |
| pFastbacHTc | green tubes, box 6 |
| pFastbac1 | purple tubes, box 6 |
| pFastbacDUAL | green tubes, box 8 |
| pFBDM | pink tubes, box 8 |
Transposition plates
Transposition plates are prepared by CSSD and are available in the second floor cold room 2S2.1
Stock solutions (1000x) for transposition plates
Link to protocol for preparation of transposition plates.
| Bluo-gal 100mg/ml | in DMSO, yellow tubes, box 9 |
| Tetracycline 10mg/ml | in ethanol, pink tubes, box 9 |
| Kanamycin 50mg/ml | aqueous, filter sterilized, purple tubes, box 9 |
| Gentamycin 7mg/ml | aqueous, filter sterilized, blue tubes, box 9 |
| IPTG 40mg/ml | aqueous, filter sterilized, green tubes, box 9 |
Primers
Primers for sequencing inserts between polyhedron promoter and terminator regions of pFastbac vectors.
| Polyhedron promoter primer (working solution in bottom of fridge 3, 5C8, stocks in freezer, box 10) |
| 5'-GGAGATAATTAAAATGATAACCATCTCGC-3' |
| 5 pmol/µl: Use 1 µl per sequencing reaction. |
| Polyhedron terminator primer (working solution in bottom of fridge 3, 5C8, stocks in freezer, box 10) |
| 5'-GCAAGTAAAACCTCTACAAATGTGG-3' |
| 5 pmol/µl: Use 1 µl per sequencing reaction. |
Primers for PCR confirmation of transposition of insert into bacmid
Link to protocol for confirmation of transposition.
| M13Fwd (box 10) |
| 5'-CCCAGTCACGACGTTGTAAAACG-3' |
| 15 pmol/µl: Use 1 µl per PCR. |
| M13Rev (box 10) |
| 5'-AGCGGATAACAATTTCACACAGG-3' |
| 15 pmol/µl: Use 1 µl per PCR. |
Competent E.coli DH10bac
Aliquots stored in -80°C Freezer A, Tray marked "Val", far left hand side of freezer, one up from bottom. Links to preparation of competent cells and transposition protocol.
Sf9 cells infected with a non-recombinant baculovirus (negative control for SDS-PAGE)
Please heat to 95°C for 3 mins and load 10 µl onto an SDS-gel. This loading provides a ladder of baculovirus infected Sf9 proteins, visible with Coomassie staining. There is generally no cross reactivity detected on blots probed with the Anti-his antibody. The gel samples are located in Box 18.