Please help in cloning gene of NDV in PC DNA 3.1 vector ?

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Narumi Uno

Dear Saddaf Razzaq

Was self-ligation occurred? Please share with us the detailed steps you have taken or are planning to take so that we can discuss further.

I think the following are some of the points that should be verified first

1. has the restriction enzyme treatment of pCDNA3.1 been done sufficiently?

2. Is the restriction enzyme treatment of insert DNA sufficient?

3. Is the dephosphorylation treatment to prevent self-ligation sufficient?

4. Is the drug selection of E. coli sufficiently performed?

Best,

Lo Tzu Yu

I think if there are a great amount of colonies without plasmid inside, you might need to check about your antibiotics