zainul at KB.USM.MY
Tue May 17 20:19:17 EST 1994
On the 16th. May 1994 my colleague, Janet Singh, wrote a question
on a problem regarding restriction enzyme Nsp-1. Up to today there are
no replies in her mailbox. Is this because she is not a subscriber to
this group or there's no answer to her problem!!
I am sending her message again. If there is a problem in sending replies
directly to her, could you please send the replies to the newsgroup.
On 16 May 1994, Janet wrote:
> Dear all, I have been using Nsp-1 restriction enzyme to cut PCR products.
> I have a control template with an Nsp-1 site which I include routinely in
> my PCR reactions to ensure the enzyme is cutting. The system was working
> perfectly until Dec 93 and since then I can't get Nsp-1 to cut. Wondering
> if I might have lost my control I tried to cut lambda DNA - nothing
> happenend!! (There should be 32 sites.) So what is wrong with Nsp-1. Is
> is by nature a sensitive enzyme?
> I have tried altering the incubation time and temperature, ordered two
> lots of news enzyme, changed the method to clean PCR products before
> enzyme incubation etc. but nothing works. Has anyone out there had
> problems with Nsp-1 or has anyone any suggestions. I'm not sure if I'm
> allowed to mention the manufacturer on e-mail but if I say the enzymes is
> supllied with x10 reaction buffer M you might have a clue!
> Please comment personally to my e-mail address since I do not subscribe
> to methods-and-reagents ( due to traffic quota on our server; one
> user subscribe to one newsgroup).
> Thank you in advance.
Zainul F. Zainuddin
Medical Microbiology & Parasitology
School of Medical Sciences,
Universiti Sains Malaysia,
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