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RNAlater stored limbal cells didn't yield any RNA, why?

RNAlater stored limbal cells didn't yield any RNA, why? - RNA Techniques Forum

RNAlater stored limbal cells didn't yield any RNA, why? - Post and discuss RNA methods and RNA science related topics. RNA extraction, cDNA synthesis, RNA EMSA, and other RNA protocols.


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Old 01-06-2012, 07:41 AM
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Default RNAlater stored limbal cells didn't yield any RNA, why?



Dear friends

I had stored the limbal epithelial cells in RNAlater (Ambion) and kept at -80 freezer. RNA extraction protocol (Qiagen) was performed on these samples after thawing. There was no RNA detected on nanodrop quantification.

Please let me know what might have gone wrong and where?? I have to finish this as soon as possible...

thanking you all for all the help and suggestion...Please help me out..

Best regards

dewoo
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Old 01-15-2012, 12:20 PM
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Default Re: RNAlater stored limbal cells didn't yield any RNA, why?

RNase contamination somewhere in the workflow? Unfortunately it's impossible to know at what stage. Be sure you work very clean and everything is RNase-free (reagents, pipet tips, tubes etc).
Another possibility is that your amount of starting material was too low. Refer to the kit protocol for the recommended amount of cells.
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Old 02-22-2012, 04:34 PM
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Default Re: RNAlater stored limbal cells didn't yield any RNA, why?

I think you should try to extract RNA from "fresh" samples and frozen ones using RNaselater and compare the result obtained from these two samples. It will be helped to figure out your problem.
Best
Sunflower_gl
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Old 02-24-2012, 02:35 PM
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Default Re: RNAlater stored limbal cells didn't yield any RNA, why?

Hi dewoo,
I have never worked with RNAlater but RNAprotect cell (Qiagen) and i have a very nice DO280/260 with the correct quantity of RNA on nanodrop.
Have you tried to extract RNA from your samples added RNAlater and from your "fresh" samples? May be it will help you to solve yr problem.
Best,
sunflower_gl
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