IUBio Biosequences .. Software .. Molbio soft .. Network News .. FTP

Problem Query

William D. Graziadei GRAZIAWD at snyplava.bitnet
Mon Nov 12 11:18:00 EST 1990

Date sent:  12-NOV-1990 11:01:52

Recently we were having problems searching for an interferon regulatory
protein clone in a lambda gt10 library. So, we decided to access the
quality of our library by screening for bovine actin cDNA (SP64-actin).
The plaques were unusual, i.e., quite small. So we decided to extract
DNA from the library using the procedure in Current Protocols in Mol. Bio.
Vol 1 (1.13.7) and hybridize the DNA blot (DNA -/+ EcoR1) to [32]P SP64-
actin. Results showed the majority of signal at 2kb (insert is 2kb) with 3
larger bands of much less intensity.However, one these signals was at ca. 50kb
for both -/+ EcoR1. Surprise and confused! Since the procedure requires the
use of DNase & RNase treatment to lysate prior to phenol extraction, could
this be the problem. Any suggestions or help would be appreciated. We are
in the process of extracting the DNA without DNase & RNase. Send responses
to GRAZIAWD at SNYPLAVA.BITNET. Thanks in advance.

        Bill :-)

|   Voice: (518) 846-7144                    | William D. Graziadei, PhD  |
|     Fax: (518) 846-7144 Ext 40             | In Vitro Cell Biology &    |
|          (518) 564-7827                    |   Biotechnology            |
|  Bitnet: GRAZIAWD at SNYPLAVA                 | SUNY Plattsburgh/Miner Inst|
|Internet: GRAZIAWD at SPLAVA.CC.PLATTSBURGH.EDU| Chazy, New York 12921      |
| EMail saves ca. 5x10e8 tree cells which would cover a std printed page. |

More information about the Methods mailing list

Send comments to us at biosci-help [At] net.bio.net