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itsron's Content

There have been 8 items by itsron (Search limited from 23-February 19)


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#174282 2 peaks for identical sequences in HRM

Posted by itsron on 09 April 2015 - 03:01 AM in PCR, RT-PCR and Real-Time PCR

Thanks, will try this!




#174276 Can I use housekeeper like -actin as endogenous control for miRNA-qPCR ?

Posted by itsron on 08 April 2015 - 10:27 PM in siRNA, microRNA and RNAi

Have you seen this link? It might help you.




#174275 2 peaks for identical sequences in HRM

Posted by itsron on 08 April 2015 - 10:11 PM in PCR, RT-PCR and Real-Time PCR

Hello,

 

I have 2 100% identical sequences but when doing an HRM, I am getting 2 different peaks! How is this even possible?

 

Any ideas?

 

Thanks!




#172244 about microrna overexpressing

Posted by itsron on 22 December 2014 - 02:46 AM in siRNA, microRNA and RNAi

Late by a month, but may be this and this will be of some help to you.




#172241 miRNA yield from human urine

Posted by itsron on 21 December 2014 - 09:12 PM in siRNA, microRNA and RNAi

It could be the kit. Qiagen is one of the top kit producers, but I have got better concentrations from other kits made by other companies too. Don't want to mention any names here.

So which one do you suggest? Otherwise it is not going to help me!




#172215 Sperm microRNA isolation

Posted by itsron on 19 December 2014 - 08:36 PM in siRNA, microRNA and RNAi

Did you try Trizol (Invitrogen) method?




#172214 miRNA yield from human urine

Posted by itsron on 19 December 2014 - 08:28 PM in siRNA, microRNA and RNAi

I recently isolated miRNA from human urine using Quigen miRNeasy serum/plasma kit (which works with urine as well) and quantified using NanoDrop. Even from freshly isolated sample, I am getting a horrible 260/280 ratios like 1.5 and 1.6. Concentration is around 8-30 ng/ul

 

My feeling is that the choice of Nanodrop is not right as miRNAs are only present in lower amounts in urine and these levels may not be detected accurately by NanoDrop. I can not imagine another reason for such a low ratio (I am pretty sure I did the experiment right).

 

Has anybody here worked with urinary miRNAs? I would like to know how much is the concentration range you are getting after isolation, and how to you quantify/qualify miRNAs?( I guess it would be Bioanalyzer).

 

Thanks mates!

 




#172213 DETERMINATION OF BASELINE FOR ANALYSIS

Posted by itsron on 19 December 2014 - 08:10 PM in Macroarray and Microarray

Since I understand you have no 'control', I wonder how was your data 'normalized'? I have seen cases of 'normalized to the median of controls' kind of cases.





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