Technique / Molecular Biology / Cloning library construction and screening


ligation reaction cosmid DNA with genomic DNA



ligation reaction cosmid DNA with genomic DNA

I would like a detailed protocol for the ligation of cosmid DNA with genomic DNA.

I have one but it ask me to set up a reaction containing a 9:1 molar ratio of vector DNA:eukaryotic DNA. I don't know how to figure out the molar concentrations. ... ...

This paper discussed cloning into cosmid vector:

Cloning of multiple copies of immunoglobulin variable kappa genes in cosmid vectors. R Cattaneo, J Gorski, and B Mach. Nucleic Acids Res. 1981 June 25; 9(12): 2777–2790.

Last update 04-Mar-2002, Rating of 0 votes.

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By cnbeatles on 25-Sep-2004
I am try to ligate a 1 kb Pst I partial digestion fragment into a 7.3 kb vector, I try 3 months, never get right plasmid. I can get colonies, but the plasmids are always strange, not what i want. Suggestions will be appreciated.
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By uma shankar on 03-Dec-2003
try following and reply plz if got sucess

vector 1ug

insert 1ug

mix both and incubate 1t 65 degree for 15 min

cool at RT for 15 min

lig buffer 1ul

T4 Ligase 1ul

--------------------

Toatal 10ul

ligate at 4 degree in refrigeretor lower most rack

best regrds

Us gautam
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By sonia on 24-Nov-2003
i am trying to prepare genomic DNA library inYEp24(7700bp) vector i am digesting vector and Genomic DNa with Bam H1 and treating my vector with CIAP.my ligation is not working .Suggestions will be appreciated
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By nitu on 11-Nov-2003
I want to clone 1.4 kb PCR fragment into 6kb vector. I am having problem doing ligation
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By Poornima on 31-Jul-2002
I am trying cloning of an eukaryotic gene into a yeast vector,I have problems with ligation,I am trying a blunt end cloning,I would be glad of i ould get some suggestions regarding the molar ratio
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By ankita srivastava on 04-Mar-2002
dear sir,

i want to know the detailed protocol to set the ligation reaction in semiliki virus plasmid
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