From owner-dros@hgmp.mrc.ac.uk Thu Oct 21 20:41:28 2004 Return-Path: X-Original-To: dros-outgoing Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with SMTP id 162D57D0CF for ; Thu, 21 Oct 2004 20:41:28 +0100 (BST) X-Original-To: dros-list@hgmp.mrc.ac.uk Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 951157D0EB for ; Thu, 21 Oct 2004 20:41:27 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6020) id A9C877D0CF; Thu, 21 Oct 2004 20:41:23 +0100 (BST) X-Original-To: bionet-drosophila@net.bio.net Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id AA1AA7D0CF for ; Thu, 21 Oct 2004 20:01:24 +0100 (BST) Received: from mx01.uchicago.edu (mx01.uchicago.edu [128.135.12.174]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 2920A7D0EB for ; Thu, 21 Oct 2004 20:01:18 +0100 (BST) Received: from churlish.uchicago.edu (churlish.uchicago.edu [128.135.12.54]) by mx01.uchicago.edu (8.12.10/8.12.9) with ESMTP id i9LIraQa027651 for ; Thu, 21 Oct 2004 13:53:36 -0500 (CDT) Received: (from nobody@localhost) by churlish.uchicago.edu (8.12.9/8.12.9) id i9LIqTVr018871 for bionet-drosophila@net.bio.net; Thu, 21 Oct 2004 13:52:29 -0500 (CDT) Received: from 128.135.150.25 ( [128.135.150.25]) as user rpbrown@nsit-imap.uchicago.edu by webmail.uchicago.edu with HTTP; Thu, 21 Oct 2004 13:52:29 -0500 Message-ID: <1098384749.4178056db4b6a@webmail.uchicago.edu> From: Rebecca Petersen Brown To: bionet-drosophila@net.bio.net Subject: website posting MIME-Version: 1.0 Content-Type: text/plain; charset=ISO-8859-1 Content-Transfer-Encoding: 8bit User-Agent: Internet Messaging Program (IMP) 3.1 X-Originating-IP: 128.135.150.25 Newsgroups: bionet.drosophila Date: Thu, 21 Oct 2004 20:41:23 +0100 (BST) Sender: owner-dros@hgmp.mrc.ac.uk Precedence: bulk Please post on your website: " I am trying to amplify 2 kb regions in the promoters of Drosophila genes and need a very high fidelity enzyme. I cannot tolerate any PCR errors. I have tried MasterAmp Extra Long Polymerase Mix from Epicentre Technologies but it is encorporates too many errors. I have been trouble shooting with Stratagene's PfuUltra Hot Start High Fidelity enzymes but I did not find appropriate sized bands even after many attempts to modify annealing temperature, and modifying amounts of DMSO, template DNA, salts, dNTPs, enzyme, or primers. Does anyone recommend a very high-fidelity enzyme for amplification of 2kb from Drosophila genomic DNA? Thanks, Becca Brown rpbrown at uchicago.edu " -- Rebecca Petersen Brown NSF Postdoctoral Fellow The University of Chicago Department of Organismal Biology & Anatomy 1027 East 57th Street Chicago, IL 60637-1508 tel. 773-834-0467 fax. 773-702-0037 email rpbrown@uchicago.edu .