cycle sequencing with 35S??
nash at biologysx.lan.nrc.ca
Wed May 19 18:59:15 EST 1993
In article <erman-190593172439 at 188.8.131.52> erman at brandeis.edu.bitnet (batu erman) writes:
>From: erman at brandeis.edu.bitnet (batu erman)
>Subject: Re: cycle sequencing with 35S??
>Date: Wed, 19 May 1993 21:28:16 GMT
>In article <C7AB34.35M at ccu.umanitoba.ca>, jrichmn at ccu.umanitoba.ca (Joy
>> Dear Netters,
>> Has anyone out there got cycle sequencing to work using 35 S? If so I
>> would like to know what kit you used and how much dsDNA you started with.
>> I would like to get away from using 32 P since we are not doing it often
>> enough to get through even the smallest amount of gamma 32 P before it
>> P.S. We have the Gibco kit but they do not recommend its use with 35 S.
>> The most important event in life is not birth or death but gastrulation.
>> Lewis Wolpert
>> jrichmn at ccu.umanitoba.ca
>Promega also says one should kinase the primers instead of incorporating
>the 35-S or 32-P into the extension products. Why is this? I cannot think
>any good reason.
'twasn't a kit... I made up the components, but NEB's Circumvent cycle
sequencing system, which uses Vent polymerase instead of Taq pol, works fine
with 35-S. Yes, there is also a kit available, (and I even used it once
<blush>, and liked it). I used the incorporation method, as opposed to the
John Nash | Email: Nash at biologysx.lan.nrc.ca.
Institute for Biological Sciences, | National Research Council of Canada,
Cell Physiology Group. | Ottawa, Ontario, Canada.
*** Disclaimer: All opinions are mine, not NRC's! ***
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