Colony Fast-Screen™ Kit (Restriction Screen)

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Last modified: 2008-07-15 14:57:15

The Colony Fast-Screen™ Kit (Restriction Screen) provides an easy and rapid method to screen the size and orientation of cloned DNA. Just like the other Colony Fast-Screen Kits (Size and PCR Screens), there is no need for overnight cultures or plasmid minipreps. The Colony Fast-Screen™ Kit Restricti-Lyse™ solution lyses cells in an environment which is not inhibitory to restriction enzyme activity. Thus, the Colony Fast-Screen™ Kit (Restriction Screen) is very convenient because it enables the use of any restriction enzyme for restriction analysis. Recombinant clone-containing E. coli colonies can be screened in 25 minutes using the standard protocol, or in as little as 10 minutes with the accelerated protocol.

Figure 1. Go from colony to gel loading in 25 minutes (or in as little as 10 minutes with the accelerated protocol) using the Colony Fast-Screen™ Kit (Restriction Screen). Pick colony into a microcentrifuge tube or microplate well. Add Restricti-Lyse™ Solution, vortex, heat for 60 seconds at 100°C. Perform restriction digestion on the entire tube of lysed cells for 15 minutes at 37°C. Load gel, electrophorese and stain.

Figure 2. Results of the standard protocol with the Colony Fast-Screen™ Kit (Restriction Screen), which goes from colony to gel loading in 25 minutes. Lane 1, molecular size marker; Lanes 2 to 8, Sca I digests of DNA from 7 colonies prepared according to the kit's standard protocol. Lanes 2, 3, and 6 show clones in orientation A; Lanes 4, 7, and 8 show clones in orientation B; and Lane 5 is a recircularized vector.

Figure 3. Results of the accelerated protocol with the Colony Fast-Screen™ Kit (Restriction Screen), which goes from colony to gel loading in 10 minutes. Lanes 1 to 7, Sca I digests of DNA from 7 colonies prepared according to the kit's accelerated protocol; Lane 8, molecular size marker. Lanes 1, 2, and 5 show clones in orientation A. Lanes 3, 6, and 7 show clones in orientation B. Lane 4 is a recircularized vector.

Last modified: 2008-05-14 16:46:46

Applications

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

Benefits

• Rapid - Perform restriction analysis on recombinant clone containing E. coli colonies in 10-25 minutes without the need for overnight cultures and plasmid preps.
• Robust - the Restriction Screen solution does not inhibit restriction enzyme activity.
• High-throughput capability - the kit is amenable to both high-throughput and routine cloning applications.
• Flexible - use the kit for clone determination as low as 100 bp.

Last modified: 2008-05-14 16:46:46

Protocols for: Colony Fast-Screen™ Kit (Restriction Screen)

(catalogue number FS0472H)

Please note: all protocols off site are the responsibility of the products supplier

Last modified: 2008-05-14 16:46:46