pGEX-MJD

Yu Wai Chen ywc at mrc-lmb.cam.ac.uk
Wed Sep 24 09:28:53 EST 1997


Dear Danilo,

I attempted once sequencing the plasmid.  There are a lot of ambiguious
bases in the sequence we obtained.  I suspect that I need to clean up
the plasmid prep before I can get a good sequence.

Now I want to digest the pSE111 and then gel-purify the mini-prep.  I
need to know some more about pSE111.

(1) What is the molecular weight of intact pSE111?
(2) I need an enzyme that cuts pSE111 but not pGEX-MJD.  I found that
HindIII does not cut pGEX-MJD.  Does it cut pSE111?  If yes, how many
pieces does it give and please give me the approx. sizes of each piece. 
If HindIII does not cut pSE111, can you suggest me another enzyme and
tell me the sizes of all the cut pieces?

Thanks a lot.

The protein sample you gave me has degraded, despite that I have kept it
at -70oC all the time until I used it yesterday.  There are a lot of low
molecular weight products but none at 70kd.  I have got some protein
from an initial trial expression but the yield is not high.

Wai
-- 
===================================================================
Yu Wai CHEN, Ph.D. ..................  email: ywc at mrc-lmb.cam.ac.uk
 Centre for Protein Engineering,              tel: 44-(1223) 402148
 MRC Centre, Cambridge  CB2 2QH, U.K.         fax: 44-(1223) 402140
 WWW homepage: http://www.mrc-cpe.cam.ac.uk/people/wai.html



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