BAC-FISH Protocol Protocol uses the BIOPRIME reaction kit from GibcoBRL to prepare biotin-labelled BAC DNA which is detected using FITC-Avidin (Vector Labs, DCS grade). Reagents from other manufacturers may work equally well but have not been tested. Includes: Labeling of BAC clones; Ethanol precipitation; Hybridization; Post-hybridisation treatment / detection.
BACs Bacterial Artificial Chromosomes Information for BACs, Bacterial Artificial Chromosomes. Includes: Existing vectors; pBAC108L; pBeloBAC11; pBACe3.6; pFW11 and F' factor; pCC1BAC; pSMART VC Vectors; pIndigoBAC-5; Cosmids; pWEB-TNCTM; SuperCos 1; pFos1; PAC vectors; BAC purification; Putting the BAC into E. coli.
DNA Isolation From BAC & PAC Clones Protocol Rapid alkaline lysis miniprep method for isolating DNA from large PAC clones. It is a modification of a standard Qiagen-Tip method that uses no organic extractions or columns. The method works very well for doing analytical restriction digests of PAC clones and can be scaled up if necessary.
Dye Protocols and Notes for Cosmid, BAC, BAC, Fosmid Templates Sequencing conditions tested for the ABI Big-Dye terminators (PE-ABI #4303150 for the 1000 reaction kit - Description: TF,KIT BTD RR-1000) with various templates. Important to quantitate all templates by agarose gel electrophoresis vs size and concentration standards and do a few tests with different template concentrations to determine the optimal conditions for your reactions. Several conditions are given.
Isolation of BAC DNA from Small-scale Cultures Protocol BAC DNAs are prepared from 5-ml cultures of BAC-transformed cells by a modification of the standard alkaline lysis method (Preparation of Plasmid DNA by Alkaline Lysis with SDS: Minipreparation). The yield typically varies between 0.1 and 0.4 µg of BAC DNA.