Home | Community | Message Board


High Mountain Compost
Please support our sponsors.

General Interest >> Science and Technology

Welcome to the Shroomery Message Board! You are experiencing a small sample of what the site has to offer. Please login or register to post messages and view our exclusive members-only content. You'll gain access to additional forums, file attachments, board customizations, encrypted private messages, and much more!

Jump to first unread post. Pages: 1
Invisiblemicro
bunbun has a gungun
Male User Gallery

Registered: 05/09/03
Posts: 7,532
Loc: Brick City
Site-Directed Mutagenesis
    #2033481 - 10/22/03 07:56 PM (13 years, 7 months ago)

O.k. -- I have to change the genome of a plasmid because one of the sites that's used for antibody production is in two different places. My boss wants me to change a G to a C in the beginning of the site (a transposon.) There's this kit, apparently, to do it, and the instructions seem pretty straight-forward, but how the fuck does this work?

It uses a thermocycler and the number of cycles, apparently, determines the type of mutations....

I looked it up and couldn't find anything. Thanks,

--
Micro


--------------------
Any research paper or book for free
(Avatar is Maxxy, a character by Mizzyam, RIP)


Post Extras: Print Post  Remind Me! Notify Moderator
OfflineMetaShroom
菌类
 User Gallery

Registered: 06/02/02
Posts: 1,462
Loc: East Anglia UK
Last seen: 9 years, 11 months
Re: Site-Directed Mutagenesis [Re: micro]
    #2035740 - 10/23/03 12:22 PM (13 years, 7 months ago)

The detail would depend on which kit you are using but basically you need two primers containing the mutation which are complimentary to the sections of plasmid either side of your mutation site. These are then extended by a polymerase leaving nicks and the original DNA is digested by something like DpnI which only targets methylated DNA. The nicks are repaired when you later tranform the mutated plasmid.


--------------------
:sleepingcow:  :penguinmonkey: :blah:

JOIN MAPS -> www.MAPS.ORG


Post Extras: Print Post  Remind Me! Notify Moderator
Invisiblemicro
bunbun has a gungun
Male User Gallery

Registered: 05/09/03
Posts: 7,532
Loc: Brick City
Re: Site-Directed Mutagenesis [Re: MetaShroom]
    #2035779 - 10/23/03 12:38 PM (13 years, 7 months ago)

Thanks -- that makes sense :wink:

BTW -- it's something like the E-Z QuickChange kit made be some company like stargene or something -- don't know exactly off the top of my head and I'm not in that lab.

--
Micro


--------------------
Any research paper or book for free
(Avatar is Maxxy, a character by Mizzyam, RIP)


Edited by micro (10/23/03 03:44 PM)


Post Extras: Print Post  Remind Me! Notify Moderator
Jump to top. Pages: 1

General Interest >> Science and Technology

Similar ThreadsPosterViewsRepliesLast post
* Comcast Sued for Blocking P2P Sites DiploidM 802 1 11/16/07 08:21 AM
by Seuss
* Can anyone confirm the validity of this PSP download site? Newbie 1,029 13 03/18/08 06:56 PM
by Cepheus
* Good *free* FTP site? bandaid 758 6 08/15/04 12:20 AM
by skunkjar
* UFO SITING OMFG!!!!
( 1 2 3 all )
CptnGarden 4,527 53 06/16/05 09:11 PM
by nightkrawler
* $13 billion nuclear fusion site gets green light Le_Canard 1,474 14 07/01/05 04:13 AM
by Diploid
* what site has the cheapest computer parts?? A3eyedfish 634 6 12/28/07 04:08 AM
by supra
* omni vs directional antenna vonhumboldt 1,230 9 10/06/08 10:16 PM
by Baby_Hitler
* Question about submitting your site to google Jalruza 1,357 8 02/20/07 11:22 AM
by Diploid

Extra information
You cannot start new topics / You cannot reply to topics
HTML is disabled / BBCode is enabled
Moderator: Lana, trendal, automan
572 topic views. 1 members, 1 guests and 1 web crawlers are browsing this forum.
[ Toggle Favorite | Print Topic | Stats ]
Search this thread:
Avalon Magic Plants
Please support our sponsors.

Copyright 1997-2017 Mind Media. Some rights reserved.

Generated in 0.026 seconds spending 0.008 seconds on 18 queries.