PCR overlap troubleshooting-HELP!

Wang haiyan hayawang at PRIDNS.SCUU.EDU.CN
Thu Mar 25 10:24:12 EST 1999


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Hello,
    I have a question regarding PCR overlap. I want to ligate two =
fragments through PCR overlap extension. The length of the two fragments =
are 800 and 1100 bp separately. The overlap region is 18 bp. I purified =
the initial PCR products from gel and do recombinant PCR. The annealing =
temp. ranges from 45-60. The product is smear and no proper 1.9 kb band =
appear. How can I do with it?
    Any suggestion will be greatly appreciated.
    Thank you

    Haiyan Wang

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<META content=3Dtext/html;charset=3Dgb2312 http-equiv=3DContent-Type>
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<BODY bgColor=3D#ffffff>
<DIV><FONT color=3D#000000 size=3D2>Hello,</FONT></DIV>
<DIV><FONT color=3D#000000 size=3D2>&nbsp;&nbsp;&nbsp; I have a question =
regarding=20
PCR overlap. I want to ligate two fragments through PCR overlap =
extension. The=20
length of the two fragments are 800 and 1100 bp separately. The overlap =
region=20
is 18 bp. I purified the initial PCR products from gel and do =
recombinant PCR.=20
The annealing temp. ranges from 45-60. The product is smear and no =
proper 1.9 kb=20
band appear. How can I do with it?</FONT></DIV>
<DIV><FONT color=3D#000000 size=3D2>&nbsp;&nbsp;&nbsp; Any suggestion =
will be=20
greatly appreciated.</FONT></DIV>
<DIV><FONT color=3D#000000 size=3D2>&nbsp;&nbsp;&nbsp; Thank =
you</FONT></DIV>
<DIV><FONT color=3D#000000 size=3D2></FONT>&nbsp;</DIV>
<DIV><FONT color=3D#000000 size=3D2>&nbsp;&nbsp;&nbsp; Haiyan=20
Wang</FONT></DIV></BODY></HTML>

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