COLPCR
Colony PCR Kit
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About This Item
UNSPSC Code:
41106300
shipped in
wet ice
storage temp.
−20°C
Application
Isolating DNA to verify the cloning insert after transformation is often time consuming. While avoiding DNA purification is not practical in many cases, it is advantageous to avoid such time-consuming steps when possible. Using the Colony PCR Kit, DNA isolation is not necessary. Take a small amount of the colony or culture, add the kit components, and proceed to PCR. Clone identification can be verified faster and more efficiently. Gel electrophoresis may not be necessary depending on primer design or other DNA detection methods available. This time-saving system is perfect for screening just a few colonies or for high throughput screening of many clones.
A dot blot method of qualitative detection may be used after PCR if a primer specific to the insert of interest is used. A PCR product will be obtained only if the colony contains the insert. Ethidium bromide is added to an aliquot of the PCR reaction after thermal cycling and will fluoresce if DNA has been amplified. JumpStart™ REDTaq ReadyMix allows for direct loading of the samples after PCR without the need of additional buffers or loading dyes. The inert dye migrates slightly faster than bromophenol blue at the same rate as a 125 base pair fragment.
A dot blot method of qualitative detection may be used after PCR if a primer specific to the insert of interest is used. A PCR product will be obtained only if the colony contains the insert. Ethidium bromide is added to an aliquot of the PCR reaction after thermal cycling and will fluoresce if DNA has been amplified. JumpStart™ REDTaq ReadyMix allows for direct loading of the samples after PCR without the need of additional buffers or loading dyes. The inert dye migrates slightly faster than bromophenol blue at the same rate as a 125 base pair fragment.
Features and Benefits
- Ultimate Flexibility. DNA purification is not necessary. Suitable for use with common various strains of E. coli, such as DH5a, JM109, and HB101 strains, as well as yeast cells of S. pombe. Amplify from as little as ~5000 copies of plasmid or ~350 colonies containing pBR322 and detect vector inserts up to 1.4 kb
- Greater Specificity & Increased Target Yield. JumpStart Taq, an antibody inactivated hot start enzyme, is designed to minimize non-specific amplification while increasing target yield & specificity
- High Tolerance. JumpStart REDTaq ReadyMix has been shown to tolerate LB concentrations up to 20% and LB Agar or Terrific Broth up to 10%.
Preparation Note
Sufficient for 100 50-μl reactions.
Legal Information
Use of this product is covered by one or more of the following US patents and corresponding patent claims outside the US: 5,789,224, 5,618,711, 6,127,155 and claims outside the US corresponding to expired US Patent No. 5,079,352. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser′s own internal research. No right under any other patent claim, no right to perform any patented method, and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser′s activities for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patents require a separate license from Roche. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.
JumpStart is a trademark of Sigma-Aldrich Co. LLC
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