Electropor. in Yeast

BRAINSL at aol.com BRAINSL at aol.com
Mon Sep 25 07:55:11 EST 1995

This email was sent to us in error.  This email must belongs to
BRIANSL at aol.com.    BRIAN is a guys NAME!

We are BRAINSL (as in the human mind)  Please take us off your email listing.
 Thank you.


ok, In a message dated 95-09-20 11:03:09 EDT, BIOSCI-REQUEST at net.bio.net

>Subj:	RE: Electropor. in Yeast
>Date:	95-09-20 11:03:09 EDT
>From:	tomasd at bdg10.niddk.nih.gov (Tomas Drgon)
>Sender:	postman at alw.nih.gov (AMDS Postmaster)
>Resent-from:	BIOSCI-REQUEST at net.bio.net
>To:	yeast at net.bio.net
>In article <cve-1509951217500001 at>, cve at ifm.mh-hannover.de
>(Christian Velten) wrote:
>> Hi everybody,
>> does anyone know a good source for the electroporation of S. cerevisiae
>> Thanks,
>> Christian
>A protocol usually comes with the cuvettes. This one came with the BRL ones:
>1. Grow cells in YEDP (2% pep, 1% YE, 2% gluc.) 50 ml. Harvest in log
>phase (5x10^7 cells/ml).
>2. Centrifuge at 2000 g 5 min 4oC. Wash cells 2x by resuspending in 25 ml
>of cold sterile water. (The goal is to decrease the conductivity)
>3. Resuspend cells in 250 ul of cold sterile water. Store on ice over
>short time. Some people even claim you can freeze it as E.coli (never
>4. Use 20 ul for 1 electroporation. Add 1 ug DNA and hold on ice for 15 min.
>5. Suspend all 21 ul between the bosses of the electroporation cuvette.
>6. Electroporator setting: 400 V, 10 uF and low ohmes.
>7. Remove cell suspension from the cuvette, add 79 ul sterile water, and
>plate the whole volume on a selective or whatever plate.
>rem:  some people recommend 1M sorbitol instead of water from step 3 and
>also in the plates.
>Tomas Drgon
>Vidno jak psoj kulky.
>(Old Slovak proverb)
>----------------------- Headers --------------------------------
>From BIOSCI-REQUEST at net.bio.net  Wed Sep 20 10:41:51 1995
>Return-Path: BIOSCI-REQUEST at net.bio.net
>Received: from OCELOT.RUTGERS.EDU (ocelot.rutgers.edu []) by
>emin01.mail.aol.com (8.6.12/8.6.12) with SMTP id KAA02939 for
><brainsl at aol.com>; Wed, 20 Sep 1995 10:41:48 -0400
>Resent-From: BIOSCI-REQUEST at net.bio.net
>Received: from mbcl.rutgers.edu by mbcl.rutgers.edu (PMDF #12194) id
> <01HVHVCFUT8WA9MG88 at mbcl.rutgers.edu>; Wed, 20 Sep 1995 10:36 EDT
>Received: from net.bio.net by mbcl.rutgers.edu (PMDF #12194) id
> <01HVHVBIIJDC9TF8TW at mbcl.rutgers.edu>; Wed, 20 Sep 1995 10:36 EDT
>Received: (from daemon at localhost) by net.bio.net (8.6.12/8.6.6) id HAA12306
> yeast-list; Wed, 20 Sep 1995 07:03:37 -0700
>Received: (from news at localhost) by net.bio.net (8.6.12/8.6.6) id HAA12300
> yeast-arpanet; Wed, 20 Sep 1995 07:03:35 -0700
>Resent-date: Wed, 20 Sep 1995 10:36 EDT
>Date: Wed, 20 Sep 1995 13:55:53 GMT
>From: tomasd at bdg10.niddk.nih.gov (Tomas Drgon)
>Subject: RE: Electropor. in Yeast
>Sender: postman at alw.nih.gov (AMDS Postmaster)
>Resent-to: brainsl at aol.com,
>        Neigeborn#m#_Lenore.mbcl_microsoft_mailserver at raven.rutgers.edu,
>        Vershon#m#_Andrew_K.mbcl_microsoft_mailserver at raven.rutgers.edu
>To: yeast at net.bio.net
>Resent-message-id: <01HVHVCFUT8WA9MG88 at mbcl.rutgers.edu>
>Message-id: <tomasd-2609561137330001 at fp5-9n.domain>
>X-Envelope-to: brainsl at aol.com
>X-VMS-To: IN%"yeast at net.bio.net"
>Nntp-Posting-Host: fp5-9n.domain

More information about the Yeast mailing list