Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Protocols and Methods Forum
Register Search Today's Posts Mark Forums Read

Protocols and Methods Forum Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


Intrinsic mutation frequency in synthetic oligoes

Intrinsic mutation frequency in synthetic oligoes - Protocols and Methods Forum

Intrinsic mutation frequency in synthetic oligoes - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 03-12-2007, 03:47 PM
peter
Guest
 
Posts: n/a
Default Intrinsic mutation frequency in synthetic oligoes



Hi netters,
I need a method to assess the mutation frequency (purity) of
ologonucleotides - 50 -60 bp long. Any Idea how to do this?

Peter

Reply With Quote
  #2  
Old 03-22-2007, 09:30 AM
newsnet customer
Guest
 
Posts: n/a
Default Intrinsic mutation frequency in synthetic oligoes

> Hi netters,

clone and sequence

St


Reply With Quote
  #3  
Old 03-22-2007, 12:01 PM
Aawara Chowdhury
Guest
 
Posts: n/a
Default Intrinsic mutation frequency in synthetic oligoes

In <1173714423.081596.43410@c51g2000cwc.googlegroups. com>,
peter <[Only registered users see links. ]> wrote:


Oligonucleotide purity is typically assessed by MALDI-TOF mass
spec, or electrospray ionization mass spec (ESI). There is a
relatively recent paper, titled "High-throughput analysis of
oligonucleotides using automated electrospray ionization mass
spectrometry", by M.E. Hail et al. that compares the two
techniques (Am. Biotech. Lab 22: 12-24).

ESI was reliable for oligonucleotides upto 120 nt in length.
MALDI-TOF was reliable up to about 50 nt.

AC
--
Email: echo 36434455860060025978157675027927670979097959886449 930P | dc
Reply With Quote
  #4  
Old 03-22-2007, 02:38 PM
peter
Guest
 
Posts: n/a
Default Intrinsic mutation frequency in synthetic oligoes

On Mar 22, 8:01 am, Aawara Chowdhury <[Only registered users see links. ]> wrote:

Thanks,
ESI and MALDI is usually provided by the supplier. I want to know
minor (less than 1%) fraction that might carry mutations. I was
thinking about sequencing, but that is quite brutal force and won't
give me results easily. Maybe I would make a construct with out of
frame AbtR and then I will clone my oligo (keeping AbtR out of frame)
and plate on antibiotic to see how many clones are resistant (e.g.
having oligoes that have 1bp deletions and repair the frame). Any
thoughts?
Peter

Reply With Quote
  #5  
Old 03-22-2007, 09:44 PM
DK
Guest
 
Posts: n/a
Default Intrinsic mutation frequency in synthetic oligoes

In article <seuMh.10918$[Only registered users see links. ]>, [Only registered users see links. ] wrote:

My non-expert understanding is that mass-spec is not particularly
(if at all) quantitative. If so, those spectra supplie by oligo
companies only detect gross impurities.

DK
Reply With Quote
Reply

Tags
frequency , intrinsic , mutation , oligoes , synthetic


Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
Plant Synthetic Biology Post Doctoral Positions Medford,June Arabidopsis and Plant Biology 0 02-04-2009 11:18 PM
Photoelectric effect is not quantized h_v_ansari@yahoo.com Physics Forum 1 10-26-2006 01:23 PM
[Drosophila] Mutation not as described Keith Maggert Drosophila Forum 0 07-25-2005 06:11 PM
Velocity is the frequency of unit displacement. hemetis@gmail.com Physics Forum 16 06-16-2005 03:46 PM


All times are GMT. The time now is 04:16 PM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.13308 seconds with 16 queries