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megaplasmid transformation/electroporation help!


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#1 izeman

izeman

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Posted 01 December 2004 - 10:23 PM

Hello ppl,

I am trying to transform a wildtype (mega)plasmid (22kb) from Pseudomonas putida to E.coli.The competent cells am using DH10B has so far given no transformants.I have tried both electroporation and CaCl transformation.

Electroporation Condition:

Elctroporation Apparatus: BIO RAD Gene Pulser 2
voltage 2.5kv
Comp. cells 40microliters
Tme constant 5.06
Cuvette Size: 0.2cm
Antibiotic selection : Ampicillin

Can anybody help me out with this please.




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