BACs Bacterial Artificial Chromosome Protocols

A Triple Color FISH Technique for Mouse Chromosome Identification Describe a set of BACs that map close to the centromeric and distal chromosomal ends for each mouse chromosome.

Amplification of Insert End Sequences from BACs Clones by Thermal Asymmetric Interlaced PCR Protocol presents the amplification of insert end sequences from bacterial artificial chromosome clones using TAIL-PCR. The amplified products are suitable as probes for chromosome walking and genome mapping and as templates for direct sequencing. The protocol has been used in rice genome studies.

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.

Isolating BAC DNA from Bacterial Cultures Using the NucleoBond System Protocol Protocol describes isolation of BAC DNA from bacterial cultures using the commercial NucleoBond system.

Isolation of BAC DNA from Large-scale Cultures Protocol The procedure for isolation of BAC DNA is scaled-up to accommodate 500-ml cultures, which, on average, yield 20-25 µg of purified BAC DNA.

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.

Preparation of BAC DNA with CONCERT High Purity Plasmid Purification System Protocol Protocol for the preparation of BAC (Bacterial Artificial Chromosome) DNA with CONCERT high purity plasmid purification system. Includes:

Purification of BAC DNA Prepared with the NucleoBond System Protocol Protocol describes purification of BAC DNA that has been prepared with the commercial NucleoBond system.

Working with Bacterial Artificial Chromosomes Protocol Protocol describes how to transform E. coli with BAC DNA by electroporation.