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Protocols
- Cracking Gel Method To Screen Bacterial Colonies (Pamela Stanley Lab Wiki, Yale University)
A cracking buffer is used to crack open the bacterium, releasing its DNA (both genomic and plasmid), which is electrophoresed on an agarose gel. The genomic DNA being Mbs in length migrates very slowly while the plasmid DNA move much faster.
http://stanxterm.aecom.yu.edu/wiki/index.php?page=...
Added: Thu Mar 26 2009, Hits: 1040, Reviews: 0 Write review Cached - Quick Screening Of Colonies For Plasmid With Insert (Pamela Stanley Lab Wiki, Yale University)
In cases where a lower efficiency of right ligation is expected (such as in case of blunt-end cloning), one may need to screen many colonies. Kit based methods can become cost or effort-intensive. The following three are alternate and cheap methods in such cases: PCR-based screening, Restriction enzyme digestion-based screening and Plasmid DNA migration-based screening
http://stanxterm.aecom.yu.edu/wiki/index.php?page=...
Added: Thu Mar 26 2009, Hits: 1682, Reviews: 0 Write review Cached -
Bacteria Storage
(Gerard R. Lazo)
Preserve E. coli strains in stab agar or glycerol for mid-term storage and lyophilize for long-term storage.
http://wheat.pw.usda.gov/~lazo/methods/lazo/bactst...
Added: Tue May 14 2002, Hits: 453, Reviews: 0 Write review Cached -
Bacterial Colony PCR
(Karl Clark)
Allows quick identification of correct clones if primers exist that allow clone identification by size or hybridization.
http://www.cbs.umn.edu/~kclark/protocols/bactPCR.h...
Added: Tue May 14 2002, Hits: 261, Reviews: 0 Write review Cached -
Screen of Recombinant
(Chen's Lab)
Four methods are introduced including clony PCR, boiling ..
http://www.biochem.ucl.ac.uk/~chen/protocols/colon...
Added: Tue May 14 2002, Hits: 1164, Reviews: 0 Write review -
Screening clone by PCR
(Herskowitz Lab)
Add colony directly into PCR tubes without plasmid preparation
http://www.sacs.ucsf.edu/home/HerskowitzLab/protoc...
Added: Tue May 14 2002, Hits: 1225, Reviews: 0 Write review -
Toothpick Plasmid Assay
(Donis Keller Lab)
The toothpick assay is used to identify bacterial colonies containing recombinant plasmids following transformation and to obtain an approximation of the plasmid insert sizes.
http://hg.wustl.edu/hdk_lab_manual/plasmid/plsmid0...
Added: Tue May 14 2002, Hits: 462, Reviews: 0 Write review Cached -
Transfer of DNA from Bacterial Colonies to Nylon Membrane or Nitrocellulose
(Donis Keller Lab)
To transfer DNA from bacterial colonies to nylon (or nitrocellulose) membrane so that a library can be screened by hybridization.
http://hg.wustl.edu/hdk_lab_manual/plasmid/plsmid0...
Added: Tue May 14 2002, Hits: 131, Reviews: 0 Write review Cached