Size fractionating before making cDNA library

Zhonglin Chai zchai at COBRA.PATH.MONASH.EDU.AU
Mon Oct 27 02:16:03 EST 1997


You may use Sephacryl S400 spun column from Pormega. I use it to eliminiate
DNA fragment smaller than about 400 bp, and routinely use it to clean my
PCR products larger than 500bp (to eliminate primers and any smaller
products) for sequencing. It works very well and you usually get pretty
high recoveray rate.

Hope this of help.

ZhongLin Chai, PhD
____________________________________________
Department of Pathology and Immunology
Monash University Medical School
Alfred Hospital
Commercial Rd, Prahran, VIC 3181, AUSTRALIA
Telephone: (61 3) 9276 2698 (lab)
           (61 3) 9276 2696 (office)
Fax:       (61 3) 9529 6484
email:     zchai at cobra.path.monash.edu.au
____________________________________________



>X-Sender: tyarovin at red.weeg.uiowa.edu
>
>This is a novice's quiestion.
>
>I am trying to make a cDNA library with Lambda ZAP II vector. I need to
>size fractionate cDNA, which was ligated to adaptors, before ligating to
>vector arms. I did not label the cDNA and have less than 300 ng of cDNA,
>that's why I cannot monitor fractions collected from Sepharose column
>neither with counter nor on a gel. I have a stupid idea to add some
>markers (Lambda or Phix 174 digests) to my sample to monitor the fractions
>on agarose and/or polyacrylamide gel.
>
>So, your critique is wellcome.
>
>Timur Yarovinsky, Ph.D., M.D.
>Oakdale Research Park, B141 MTF
>2501 Crosspark Road
>Coralville IA 52241-8802
>
>tel.    (319) 335-4608
>fax     (319) 335-4347
>e-mail: timur-yarovinsky at uiowa.edu
>





More information about the Methods mailing list