sequencing strategy 2

Ari-Matti Saren amsaren at operoni.helsinki.fi
Thu Jun 20 05:02:16 EST 1996


In article <4q6b5m$i9u at net.bio.net> jea at ukrv.de writes:
>From: jea at ukrv.de
>Subject: sequencing strategy 2
>Date: 18 Jun 1996 06:32:38 -0700

>I want to know if someone can help me to decide which sequencing strategy
>is the best one. I want to start sequencing on a ABI377. I want to
>sequence PCR-Products after cloning them in pUC18 (sureClone Kit,
>Pharmacia). 

Do you really need them cloned, or do you just want the sequence? If you 
just need the sequence it's best not clone them (the possibility of PCR 
errors, you see), but rather cycle sequence the PCR fragments straight.

> My question is: is it better to sequence plasmid DNA with the
> cycle sequencing Kit or is it better to do solid phase sequencing
> (Dynabeads) and use Sequenase? 

Cycle sequencing is easier and faster. In my experience using T7 is only 
really worth it if you are searching for heterozygotes or something in that 
vein.

Ari-Matti
--
***********************   Ari-Matti Saren   **************************
*  WORK: Institute of Biotechnology, DNA Synthesis & Sequencing Lab. *
* Biokeskus 1A, PL 56 (Viikinkaari 9), Finland, Tel +358 0 708 59411 *
*    NET: Ari-Matti.Saren at Helsinki.fi, http://anime.jyu.fi/~saren    *
*         HOME: Ohjaajantie 18 A 2, 00400 Helsinki, Finland          *
**************    Shikisoku - Zeku, Kusoku - Zeshiki   ***************





More information about the Autoseq mailing list