Colony Fast-Screen™ Kit (Restriction Screen)

Applications

• Identification of desired clones (size and orientation) from a cloning experiment or in a library.
• Releasing DNA for restriction analysis.

The Colony Fast-Screen™ Kit (Restriction Screen) provides an easy and rapid method to screen the size and orientation of cloned DNA by restriction digests, without overnight cultures or plasmid minipreps (Fig. 1). The Colony Fast-Screen Kit Restricti-Lyse™ solution lyses cells in an environment which is not inhibitory to restriction enzyme activity. Thus, the kit enables the use of any restriction enzyme for restriction analysis. E. coli recombinant colonies can be screened in 25 minutes using the standard protocol (Fig. 2), or in as little as 10 minutes with the accelerated protocol (Fig. 3).

Benefits

• Perform restriction analysis on recombinant clone containing E. coli colonies in 10-25 minutes without the need for overnight cultures and plasmid preps.
• The Restriction Screen solution does not inhibit restriction enzyme activity.
• Amenable to high-throughput applications.
• Can detect inserts as small as 100 bp.

Notes:

1. The accelerated protocol involves heating the Restricti-Lyse™ Solution for 10 seconds instead of 60 seconds and incubation of the restriction digest for 5 minutes instead of 15 minutes. Restriction bands stain less intensely than when using the standard protocol, but in most cases yield a clearly discernible pattern.
2. We do not recommend using the Colony Fast-Screen Kit (Restriction Screen) with single-copy-number clones.

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