need suggestions to make Xi-clone kit working - asking foe help (Sep/11/2005 )

Hi,
I am using Xi-CloneTM PCR cloning kit from gene therapy to clone my PCR product in phCMV1 vector.
My PCR product is 2.7 Kb, and the yield is not too low. But unfortunately, I get no colony after transformation (I followed the protocol on the manual).

Anyone with experiences using this kit can give me a suggestion? Thanks in advance

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Hi,

I am from GTS tech support, and I came across your question by chance.

My suggestions would be:

1. Use use our SmartCells competent E. coli (or other library efficiency DH5alpha comp cells) if you are not already.

2. Try increasing the amount of PCR fragments used by 1 mg increments. Alternatively, you can try increasing the amount of vector two-fold to help improve the efficiency of the cloning reaction.

Please call me at 888-428-0558, Ext. 6125 if you are still having trouble.

Mike V.

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