Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Animal and Molecular Model Systems > Zebrafish Forum
Register Search Today's Posts Mark Forums Read

Zebrafish Forum Zebrafish Danio Rerio Forum. Discuss zebrafish, aquarium culture and molecular biology lab specimens.


Determining morpholino concentration

Determining morpholino concentration - Zebrafish Forum

Determining morpholino concentration - Zebrafish Danio Rerio Forum. Discuss zebrafish, aquarium culture and molecular biology lab specimens.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 07-31-2012, 05:00 PM
Pipette Filler
Points: 177, Level: 3 Points: 177, Level: 3 Points: 177, Level: 3
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jul 2012
Posts: 4
Thanks: 0
Thanked 0 Times in 0 Posts
Question Determining morpholino concentration



Hi all,

I am injecting some fish and seeing a phenotype but I am not sure of the concentration my fish are receiving (ng total). Can you guys help me figure it out?

I diluted my stock to 1mM and inject the fish with 4.6nL of 0.4mM solution. The MW of the MOs are 8474 and 8458
Reply With Quote
  #2  
Old 08-01-2012, 05:15 PM
Jon Moulton's Avatar
Post-Doc
Points: 8,218, Level: 63 Points: 8,218, Level: 63 Points: 8,218, Level: 63
Activity: 99% Activity: 99% Activity: 99%
 
Join Date: May 2007
Location: Corvallis, OR, USA
Posts: 240
Thanks: 3
Thanked 15 Times in 15 Posts
Default Re: Determining morpholino concentration

0.4 mM = 0.0004 mole/l
4.6 nl = 4.6E-09 l
mole/l * l = mole
1.84E-12 mole


For 8474 g/mole
mole * g/mole = g
1.55922E-08 g = 16 ng


For 8458 g/mole
mole * g/mole = g
1.55627E-08 g = 16 ng

I think most reviewers would call that a high dose of Morpholino, high enough that the observed phenotype might be caused by off-target RNA interactions. Have you confirmed your concentration of Morpholino in the stock solution by UV spec?
[Only registered users see links. ]
If some of the Morpholino has come out of solution, then you could be delivering a lower dose than you think -- in your case, that would be good as you would be getting a phenotype at a lower dose than you thought (but you need to measure the actual concentration of the solution to ascertain whether or not that is the situation). Be sure to heat the oligos prior to aliquoting, dilution or injecting; you want all of the oligo to be uncomplexed in solution, available as active antisense.
__________________
Jon D. Moulton, Ph.D.
Gene Tools, LLC
[Only registered users see links. ]
Reply With Quote
  #3  
Old 08-02-2012, 05:23 PM
Pipette Filler
Points: 177, Level: 3 Points: 177, Level: 3 Points: 177, Level: 3
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Jul 2012
Posts: 4
Thanks: 0
Thanked 0 Times in 0 Posts
Default Re: Determining morpholino concentration

Thank you! I thought I was going crazy when I was getting numbers that high...

I'll spec them right away!
Reply With Quote
Reply

Tags
concentration , determining , morpholino


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
Need Help Determining Concentration of DNA from Gel Electrophoresis BrianGumble Agarose Gel Electrophoresis Forum 2 02-05-2010 04:05 PM
Low concentration miniprep Utsugi, Heidi K Protocols and Methods Forum 0 01-15-2009 06:16 PM
[Fwd: RE: [Zbrafish] About morpholino heating before injection] Jon D.Moulton Zebrafish Forum 0 01-14-2009 08:19 PM
Morpholino injection concentrations and carriers? Jon D.Moulton Zebrafish Forum 2 05-26-2007 07:22 PM
[Zbrafish] Low efficacy of splice-blocking morpholino Burdine, Rebecca D Zebrafish Forum 0 03-12-2007 07:42 PM


All times are GMT. The time now is 09:15 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 0.12947 seconds with 16 queries