PROBLEMS (Nsp-1)

Zainul Zain zainul at KB.USM.MY
Tue May 17 20:19:17 EST 1994



Dear netters,

	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. 

Thank you.

 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,
Kelantan, Malaysia.




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