Riboprobe background hybridisation

Andre Hamel hamel at cc.umanitoba.ca
Thu Oct 20 12:22:24 EST 1994


Greetings Klaus

Yes, Stratagene's Bluscript vectors contain sequences associated with the
Not I site in the MCS. If one MUST use this vector for making riboprobes, 'tis
strongly recommended that the region between Sac II and Sma I be removed
from the vector vector before use.

This region binds STRONGLY to rRNAs ... thus, riboprobes used will give
unacceptably high background in in situ and northerns.  Thanks to our good
friend & consumate scientist Ingo Schroedter (U of Manitoba, Physiology
Dept) for revealing this info to us a few years ago ... he discovered
this the hard way (months of work) ... he's our in situ guru! :^)

cheers t'y'all
                            ********************
Andre Hamel                                    email: hamel at cc.umanitoba.ca
Manitoba Government				tel: 204/945-7630
Department of Agriculture  			FAX: 204/945-8062
Veterinary Services Branch          
Winnipeg, Manitoba, CANADA 
			  ********************

In <9410192338.AA22415 at cscgpo.anu.edu.au> Klaus.Matthaei at ANU.EDU.AU writes:

>Some time ago (a year or so) on the net there was some discussion about a
>certain cloning vector that had a multiple cloning site that gave high
>background when used for Riboprobe production.  Apparently part of the MCS
>sequence hybridises to ribosomal RNA.  At the time I kept a copy but have
>now unconveniantly lost it.  My feeling is that it was Bluescript II but I
>cannot be certain.  I am transferring a sequence to a new vector for
>riboprobe production and don't want to have this problem if it can be
>avoided. >Does anyone remember? give advice?  >Cheers, Klaus



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