Applications

• Identification of desired clones in a library.
• Determining the orientation of cloned inserts.
• PCR of bacterial genomic DNA.

The Colony Fast-Screen™ Kit (PCR Screen) provides a rapid method for preparing clones for screening by PCR. Using the Colony Fast-Screen Kit (PCR Screen) there is no need to grow cultures or purify DNA prior to PCR (Fig. 1). The kit can be used with all standard E. coli hosts and all cloning vectors. Thermostable polymerase and PCR primers are not provided.

Benefits

• Identify desired clones by PCR without the need to grow cultures or purify DNA (Fig. 2).
• Enables PCR screening of high-copy clones, single-copy clones, and bacterial genomic DNA.
• Amenable to high-throughput applications.
• DNA preparation process does not interfere with pre-established PCR conditions.
• Enables both long and short PCR amplifications and can be used with all standard E. coli host strains.

Figure 1. Schematic overview of the protocol for the Colony Fast- Screen™ Kit (PCR Screen).

Figure 2. The Colony Fast-Screen™ Kit (PCR Screen) enables screening of both high-copy and single-copy clones by PCR without the need for cultures and DNA purifications. Colonies containing a 5-kb lambda fragment cloned into the CopyControl™ pCC1™ vector and grown on plates at either single-copy or high-copy number were picked and processed using the Colony Fast-Screen Kit (PCR Screen). PCR was performed using primers homologous to the ends of the cloning vector. Lane 1, DNA size marker; lane 2, E. coli DNA (negative control); lane 3, single-copy clone; lane 4, high-copy clone; lane 5, PCR of 1 ng of purified DNA.