Compressed lanes--too many salts?

mlassner at ccmail.calgene.com mlassner at ccmail.calgene.com
Wed Feb 19 16:45:15 EST 1997


We use a cheaper alternative to centri-sep.  They are empty columns made 
by
Axygen, and available from USA Scientific.  They cost about $1.00 each, 
but they
are reusable many times.  we fill the columns with sephadex G-50. 
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Subject: Compressed lanes--too many salts?
From:    G.Harrison at net.bio.net, caribou at u.washington.edu at Internet_Mail
Date:    2/19/97  12:56 PM

To: autoseq at net.bio.net
Re: compressed lanes--too many salts?

Hi all.  

I've often had a problem with lanes that become compressed at the
beginning of the sequence, where the unincorporated dye terminators run.
Consequently, the lanes blur together at the bottom of the gel picture and
the computer becomes extremely confused as to which blur to assign a lane
number. 

I assume this comes from salts and stuff that are not properly cleaned up
after the thermocycling.  I notice this with samples that are EtOH ppt but
don't remember if this also occurs with centri-sep purified samples.  

Is my hypothesis correct?  Must I tell my users to buy those expensive but
useful centri-sep columns?  What is that new EtOH protocol that is floting
around (I tried to access it from an archived message, but the address
didn't work properly).  Other comments?

Replies are much appreciated,
Grace (HHMI, University of Washington)






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