High 260/230 ratio for DNA |
|
Would you like to save this topic, event, protocol or job so you can find it again easily?
Just click the "Save to My Lab Drawer" link and the item will be saved in the My Lab Drawer section of your bench space.
Available to members only. Please log in or register for your free account now.
|
|
I purified my DNA samples form whole blood using QIAamp Blood Maxi Kit (Spin Protocol). Using Nanodrop, my 260/280 ratio was ok (around 1.8), but 260/230 ratio in some samples was as high as 8! I know that ideal 260/230 ratio is around 2, and if it is lower that probably means contamination with organic solvents. Does anybody knows what high ratio means?
|
Replies
|
|
|
Posted By Shampa
on 7/3/2009 2:58 AM
|
|
You are right it should be aroung 1.5 ideally. What I know from my previous experience is that the high ratio indicates high salt concentration in your sample. Please try to reduce it.
|
|
|
|
Posted By Jiten
on 7/3/2009 16:41 PM
|
|
I may be wrong also. Shouldn't we expect the A260 / A230 ratio to be low if the salt concentration is high in the samples? I have experienced this problem and I have not been able to sort out what the error happening with similar A260 / A230 ratios. I have got even once at 18.
|
|
|
|
Posted By sps
on 7/3/2009 3:30 AM
|
|
I would advice diluting your sample in 10mM Tris-HCl at pH 7.4 but not water. One more thing, low ratios may be due to phenol contamination. Check it out!!! High ratios might come from salts in yoour samples. I never saw ratios as high as indicated here. But if you used Trizol, maybe try doing clean up using a column spin methods and try again. Wish you good luck.
|
|
|
|
Posted By Monu
on 7/3/2009 3:54 AM
|
|
I also suspect the high salts conc is the main reason behind it. But you should check spec instrument functioning also.
|
|
|
|
Posted By LBurger
on 11/9/2009 10:18 AM
|
|
Wouldn't a very high 260/230 ratio indicate a low (in in the case of some samples I just OD'd zero) absorbance at A230. Personally, I'm more concerned with contamination issues when I have significant absorbance at A230 indicating phenol, salt, etc (and low 260/230) than when it's too high. From what I've read anything over 2 is good.
Low A230, could also be related to how dilute your sample is; if the A260 is low (say 0.05), then it may be difficult to accurately estimate A230 resulting in over inflated 260/230 ratios.
I'd remeasure the samples more concentrated, if possible, and see if you can get a better A230 reading. I like to shoot for an A260 of ~0.1. However, as I said above I think a higher ratio (low A230) is better than low.
|
|
|
|
After concentrating my samples (using ethanol precipitation after adition of 3M sodium acetate) and re-measuring the concentration, my A260/230 ratio was normal! I suppose the reason for the high ratio could be high initial salt concentration, but whatever it was it is gone after this procedure 
|
|
|
|
Posted By Susie Q
on 12/13/2009 16:56 PM
|
|
I`m working with AFLPs, so I need a nice DNA that could be completely digested. I have seen that DNAs with low 260/230 ratios don`t have fully digestion... it is logical... but also DNAs with High 260/230 ratios (more than 4) just parcially digest. I am wondering myself why this could be happening????
|
|
As a Scientist Solutions member, you are able to register a positive vote for any topic which you believe is useful and relevant to our board or any reply which you believe is especially well worded and helpful.
By participating in the voting, you will be helping to identify the best topics & replies on the board.
You may vote once for any one post, and you may not vote for your own posts.
A post (topic or reply) will earn one "thumbs up" icon for every 10 votes received (up to 3 thumbs up), and the person who made the post will also earn two bonus points.
learn more about member points.
|
Become a member & join our community (It's easy & free)
Already a member? Please log in
|
Life Technologies
|
|
Life Technologies
|
|
Eppendorf
|
|
9/1/2010
|
|
8/3/2010
|
|
1/11/2010
|
|
8/7/2010
|
|
8/6/2010
|
|
8/5/2010
|
|
|