From owner-chlamy@hgmp.mrc.ac.uk Fri Aug 16 16:53:18 2002 Return-Path: Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with SMTP id DEDF617BD8 for ; Fri, 16 Aug 2002 16:53:14 +0100 (BST) Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id A993717B66 for ; Fri, 16 Aug 2002 16:53:08 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6058) id 51B7317BD6; Fri, 16 Aug 2002 16:53:03 +0100 (BST) Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id A1C1D17BE8 for ; Thu, 15 Aug 2002 17:09:55 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 60001) id CB1E817BE2; Thu, 15 Aug 2002 17:09:53 +0100 (BST) To: bionet-chlamydomonas@net.bio.net From: fleaghost@hotmail.com ("Andy Vaughan") Newsgroups: bionet.chlamydomonas Subject: Biolistic Nuclear transformation Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Message-ID: Mime-Version: 1.0 Content-Type: text/plain; format=flowed X-Received: from localhost (localhost [127.0.0.1]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 49BAB17B91 for ; Thu, 15 Aug 2002 17:09:51 +0100 (BST) X-Received: from hotmail.com (f185.pav1.hotmail.com [64.4.31.185]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 3BCBB17B96 for ; Thu, 15 Aug 2002 17:09:49 +0100 (BST) X-Received: from mail pickup service by hotmail.com with Microsoft SMTPSVC; Thu, 15 Aug 2002 09:09:47 -0700 X-Received: from 129.93.134.24 by pv1fd.pav1.hotmail.msn.com with HTTP; Thu, 15 Aug 2002 16:09:47 GMT X-Originating-IP: [129.93.134.24] X-To: chlamy@net.bio.net X-OriginalArrivalTime: 15 Aug 2002 16:09:47.0898 (UTC) FILETIME=[2D709DA0:01C24476] Date: Fri, 16 Aug 2002 16:53:03 +0100 (BST) X-Razor-id: b8b51883873b5b74480db6011f6bf8689fd84ade Sender: owner-chlamy@hgmp.mrc.ac.uk Precedence: bulk Hello all, I am attempting some experiments in which I need to do nuclear transformation with the PDS-1000/He gene gun of chlamy. I have done a large amount of shooting, but have had no good results. I've tried shooting on cells imbedded in agar and agarose, cells actually grown up on the plates, and cells on nylon membranes (the best candidate thus far), but have had no success. I've also tried a variety of pressure disks and target locations, macrocarrier locations, and microparticle preparation methods. I have tried both the pJD67 (arg plasmid) and pSP124s (ble plasmid), but have had no discernable transformants with either. I'm starting to get pretty desperate, so any help would be greatly appreciated. Thanks. Sincerely, Andy Vaughan fleaghost@hotmail.com _________________________________________________________________ Chat with friends online, try MSN Messenger: http://messenger.msn.com --- .