Problem with pART27 vector

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Vinny
Vinny's picture
Problem with pART27 vector

I have a problem with cloning pHannibal cassette in pART27....I cut phannibal cassette and open pART27 with NotI and seems to be OK....after i treat the pART27 with CIAP, extract all from gel...ligate and cloning the ligation in E.coli DH5alfa...after i select on antibiotic Kanamicin 50ug/ml...but not works....There's anyone that can help me????

Tony Rook
Tony Rook's picture
Vinny:

Vinny:

In the past, I have had problems with my antibiotic concentration being too high during selection. You may want to try and select at a lower concentrations such as 25 ug/ml.

Rock
Rock's picture
Vinny wrote:I have a problem

Vinny wrote:

I have a problem with cloning pHannibal cassette in pART27....I cut phannibal cassette and open pART27 with NotI and seems to be OK....after i treat the pART27 with CIAP, extract all from gel...ligate and cloning the ligation in E.coli DH5alfa...after i select on antibiotic Kanamicin 50ug/ml...but not works....There's anyone that can help me????
rkrl001
rkrl001's picture
Hello, will you please kindly

Hello, will you please kindly provide me pART27 vector, in my research, I really, really need it. Thank you.

The FFM
The FFM's picture
 The map can be found in this

 The map can be found in this reference

Gleave AP: A versatile binary vector system with a T-DNA organizational structure conducive to efficient integration of cloned DNA into the plant genome.
Plant Mol Biol 1992, 20:1203-1207. PubMed Abstract | Publisher Full Text 
Return to text

http://www.springerlink.com/content/w350542n1224856j/fulltext.pdf

Cao Hua
Cao Hua's picture
 Do you have the complete

 Do you have the complete sequece of the binary vector pART 27 ?  I want to change the restriction enzyme sites on it.  Thank you very much.