Technique / Molecular Biology / PCR / Inverse PCR


Inverse PCR



Inverse PCR

I am trying to amplify a restriction fragment
from genomic DNA, where I know the sequence of half of the fragment. I am
trying to amply the other half. I ligated the fragment to form circles and
trying to amplify ... ...

Last update 02-May-2002, Rating of 2 votes.

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By Nina on 17-Sep-2008
I know three quarters of my gene and used IPCR to get the rest of the flanking regions. I thought the PCR worked when I got a signal with primers that were originally pointing outwards, hence indicating that the re-ligation of the fragment containing the gene had worked. However, sequencing with the forward M13 only had given a different blast result although my forward Primer is present 100%. Please advice on this as the seq immediately next to the primer is different from my known sement from which i designed my primers for the inverse PCR procedure. What went wrong?
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By manju on 16-Oct-2005
respected sir/madam
ihave a known sequence of about 500bp RGA sequence & want extend on either sides to clone full length sequence, kindly suggest me to how to go ahead.
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By julianne on 02-May-2002
I want to find the unknow gene sequence , and the sequence was from virus(herpesvirus)

I want to trying inverse PCR . it have a fragment about 540bp was known, if i used this known fragment(540bp) to amplification

about 3~4kbp by inverse PCR , how can I do?

and how to design the experiment ?please help me ,and thank you very much!
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