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cloning problem for expression vector

cloning problem for expression vector - Protocols and Methods Forum

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Old 10-06-2004, 01:45 AM
shazdefaz@yahoo.com
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Default cloning problem for expression vector




hello,

i'm working on a plasmid pMG36e and pNZ8048 plasmid vectors for lactococcus lactis. I'm trying to clone an insert into them but to no avail.

generally i clone the insert and transform into e.coli (as done in revision review) to amplified the plasmid but no result.
i've check the RE used for digestion and it is OK.
and i've tried a lot of ligase from different product but no result.

can anyone please help me with this? i'm already at it for months now.

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Old 10-08-2004, 12:13 PM
Wolfgang Schechinger
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Default cloning problem for expression vector

Without knowing any details about the plasmids (who does?)

Do they have a chance to replicate in E. coli as they're apparently
designed for L.l. spec? (Ori, appropriate antibiotic resistance, vector
compatible with strain)

Do you see ligation products when you run an aliquot of the lig rxn on agel?
which enzmes do you use?

Please give us more details!

Best of luck and a nice weekend!

Wo

At 01:45 06.10.2004 -0000, you wrote:
lactococcus lactis. I'm trying to clone an insert into them but to no avail.
revision review) to amplified the plasmid but no result.
egin=0
Wolfgang Schechinger, PhD (Dr. rer. nat.)
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