Protocols
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Alkaline Lysis Minipreps of Plasmid or Cosmid DNA
(Donis Keller Lab)
Plasmid or cosmid containing E. coli cells are grown and lysed with alkali. The cell debris and chromosomal DNA is precipitated with SDS and potassium acetate. After pelleting the debris the plasmid/cosmid DNA is precipitated from the supernatant with isopropanol and the DNA resuspended in TE. The expected yields from a 5 ml culture are 5-10 ug for plasmids 2-5 ug for cosmids. The procedure can be scaled up many fold.
http://hg.wustl.edu/hdk_lab_manual/plasmid/plsmid0...
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Cosmid Cloning: Cell preparation, DNA packaging, and Cell Transfection
(NWFSC)
http://micro.nwfsc.noaa.gov/protocols/cosmid_packa...
Added: Tue May 14 2002, Hits: 1540, Reviews: 0 Write review -
Cosmid Preparation
(DNA Sequencing Facility, University of Utah Health Sciences)
Detailed protocol for preparing cosmid DNA.
http://www.hci.utah.edu/groups/sequencing/cosmid.h...
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Large Scale Plasmid, Cosmid, BAC, PAC, and Fosmid DNA Isolation by a Cleared Lysate Method Followed
(Roe Lab)
http://www.genome.ou.edu/DblAcetateProcV3.html
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Plasmid or Cosmid DNA Miniprep
(Chun-Ming Liu)
http://www.plant.dlo.nl/projects/hybtech/Liu/pcmin...
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Preparation of Cosmid DNA from 50 ml Cultures
(Donis Keller Lab)
Cosmid vectors containing foreign DNA inserts are known to replicate more slowly than small plasmids in culture. One of the reasons is believed to be the capacity of the cosmids to take larger insert DNAs (up to 45 kb) causing an increased DNA replication time. Routine plasmid miniprep protocols using 5 ml overnight cultures yield very little cosmid DNA. The following protocol is a scaled-up version of the 5 ml plasmid miniprep protocol with yields ranging from 5 ug to 10 ug of cosmid DNA. This protocol is particularly helpful for obtaining higher yields of the HTY cosmids.
http://hg.wustl.edu/hdk_lab_manual/plasmid/plsmid1...
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Sequencing off Cosmid, BAC, PAC, and Fosmid Templates with ABI Big Dye Terminators
(Roe Lab)
http://www.genome.ou.edu/big_dyes_cos_bac_pac_fos....
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Transformation of Plasmids/Cosmids into E. coli
(Donis Keller Lab)
To utilize competent E. coli bacteria to replicate a specific DNA fragment. Three methods are presented transformation of intact plasmids/cosmids ,transformation of ligation mixes and color selectability procedures.
http://hg.wustl.edu/hdk_lab_manual/plasmid/plsmid0...
Added: Tue May 14 2002, Hits: 2022, Reviews: 0 Write review Cached