Dear Sir,1. I found a contradiction in the paper that it is...
Dear Sir,
1. I found a contradiction in the paper that it is mentioned to take 680 mM of Na2hpo4 9.65g for 1 L solution. I will be highly obiliged if you could please make me understand if the final conc. (680mM) is correct or the weigh? (because it should be 96.5 g for 1L if the fina Read more Read less

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Maged Zeineldin Author Answered Dec 17, 2018
Molecular Biosciences Department, University of Kansas, USA
Thanks for the questions!
You can use either (Xg) or rpm.
I have not run the RNA on a bio-analyzer to give you the RIN. However, I ran it on agarose gel and it looked good to me.
Good luck!
and if you could let me know the RIN of this protocol
Hi, sorry to disturb you one more time, at the end of the procedure there is the centrifuge, in the paper you mention both 1000 g and 1000 rpm for 10 minutes, may be if could please tell me its 1000 rcf (g) or 1000 rpm for 10 minutes?
Maged Zeineldin Author Answered Nov 23, 2018
Molecular Biosciences Department, University of Kansas, USA
I assume this was because using the higher concentration. I believe when using the correct concentration of Na Phosphate you will get good amount of RNA. Small intestinal pieces yield more RNA than the colon but I used to get al least 20-ug from the colon as well. However, I have not used Qiagen for RNA extraction in this protocol. I used to use Trizol https://www.thermofisher.com/order/catalog/product/15596026
or TriZol followed by DirectZol for RNA extraction
https://www.zymoresearch.com/direct-zol-isolation-kits
Hi, Sorry for bothering you one more time, I am isolating the RNA with QIAGEN kit, only from large intestine, Last time i barely got any RNA. Could it be beacuse i used a higher concentration of NA2Hpo4? I m guessing you are also isolating RNA seperately from small and large intestine , and for each you are getting minimun 20 ug yield ? Please if you could suggest anything
Maged Zeineldin Author Answered Nov 23, 2018
Molecular Biosciences Department, University of Kansas, USA
Thanks for the questions!
The weight is the correct value (9.65 g/L) and you should not see any precipitate with this concentration.
For the RNA yeild, you should be able to get at least 20-ug RNA by following this protocol.
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