Do you have to YES!!

Tony Hodge tph at mrc-lmb.cam.ac.uk
Thu Apr 7 05:30:56 EST 1994


Subject: Do you have to sequence both strands?
From: John Gosink, gosink at u.washington.edu
Date: Sat, 02 Apr 1994 17:49:55 -0800
In article <gosink-020494174955 at microb3.biostat.washington.edu> John
Gosink, gosink at u.washington.edu writes:
>Hi,
>I was having a discussion with a person down the hall.  They claim
>that there is _no_ regulation that says you have to sequence both
strands
>of a length of DNA (they are working with cloned PCR products) for
>submission to Genbank and/or a referreed paper.  Do you have any
references
>on this subject?
>
>			                              -John
>
>P.S.  They read a single strand, but make it a point to read each
gel on
>two different occasions and/or by two different people.

Various comments from other contributors omitted


Has sequencing become too easy??  Obviously not, we all welcome the
advances made over the past years but I still prefer to do Shotgun
Sequencing which guarantees reads on both strands with multiple
redundancy - especially for those 'never sequenced before' clones. 
I have come across at least one published sequence which I know must
be wrong due to the nature of the protein product which has a highly
ordered structure (alpha helical coiled coil); lo and behold when I
sequence it compression rears it's ugly head, immediately apparent
from the mismatch between strands.

So, come on do the job properly. If you use synthetic primers make
sure you do both strands and at a reasonable level of redundancy,
the same goes for serial deletion methods, do it in both directions.


Tony


.
Tony P Hodge
Structural Studies Division
Medical Research Council Laboratory of Molecular Biology
Hills Road
Cambridge
CB2  2QH
UK

Phone (0223)  402260

Fax     (0223)  213556



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