Berkeley Drosophila Genome Project
Important: BDGP will conclude operations on January 1, 2026, and fruitfly.org will be retired soon after.
Please download and self-host the splice-site and promoter prediction tools from
github.com/bdgp/bdgp-splice-promoter-tools.
BDGP Resources
PCR Amplification of cDNAs fromBacterial Cultures: DGC/pOT2
Monica Moore Lab, UCSF Ernest Gallo Research Center
For Drosophila DGC set in pOT2 vector using Bacterial template:
-
transfer 5ul bacterial culture to a PCR plate.
-
Mix and aliquot the following cocktail:
| 5ul | Bacterial template |
| 2.5ul | DNTP mix (10mM each, Perkin Elmer) |
| 10ul | MgCl2, 25mM (Perkin Elmer) |
| 10ul | 10x PCR buffer |
| 0.30ul | Amplitaq gold enzyme (5U/ul, Perkin Elmer) |
| 0.025 | Pfu Turbo DNA polymerase |
| 0.05ul | Primer 1 500pmol/ul |
| 0.05ul | Primer 2 500pmol/ul |
| 72.075 DdH2O | |
| 100ul | Total |
-
Spin the plate briefly to remove any air bubbles. Use an MJ thermocycler for the following cycle conditions:
95C 10min. Denaturation 95C 45sec. Melting 64C 45sec. Annealing 72C 7min. Extension 72C 8min. Final Extension 4C forever
Cycle 35 times. Slow cool from final extension at 72C to final hold at 4C at a ramping rate of 0.1C/sec.
Vector Primer Name Sequence
POT2a OTf AATGCAGGTTAACCTGGCTTATCG
POT2a OTr AACGCGGCTACAATTAATACATAACC
pBS ESTf GAAACAGCTATGACCATGATTACGCC
pBS ESTr CGGCCAGTGAATTGTAATACGACTC
Order oligos from Operon at a 1umol scale, HPLC purified and shipped lyophilized rev. 8.23.00