Toothpick™-PCR is an extension of Toothpick™ and allows for the rapid release of plasmids from transformed bacteria for screening by polymerase chain reaction (PCR). There is no requirement for growing bacteria, performing "minipreps" or purifying the plasmid DNA. Add your colony directly to the supplied Toothpick™ solution, heat and then transfer 1µl directly to a PCR reaction to screen for your plasmid. Following PCR, add the supplied GLOW™ Loading Dye and run on an agarose gel. No need to stain with ethidium bromide, simply place gel on UV box and view. Glow Dyes™ have been specifically formulated for nucleic acid electrophoresis and contain an optimized concentration of buffer agents, tracking dyes and ethidium bromide. These loading dyes reduce exposure and many of the problems associated with ethidium bromide use and disposal.
Figure: Screening of Plasmids with Toothpick™-PCR.Four transformed bacterial colonies were picked, resuspended in 15µl Toothpick™ solution and boiled for 5 minutes.1µl was added to a 50µl standard PCR reaction and the resulting PCR products were resolved on an agarose gel.Lanes 1 and 2 show the correct plasmid.
Features
- Simple protocol, as easy as picking a colony.
- 5 minute protocol.
- No overnight cultures required.
- Enough reagent to screen 300 colonies by PCR.
- Supplied with GLOW™ Loading Dye to minimize ethidium bromide use.
Applications
- For the rapid screening of colonies using PCR analysis.