Bhoomika Sharma

9 Questions 11 Answers 0 Followers

Questions related from Bhoomika Sharma

Problem with PCR amplification?

i tried gradient PCR for a new set of primers and got the amplification. However, when i tried running the same set of primers on a single annealing temperature, it showed no or weak...

04 November 2016 6,640 13 View

Can anyone tell how to troubleshoot no amplification in PCR?

I am trying to amplify my genes of interest.Though I got the amplified product of the desired size on my first attempt , i am not able to get the amplification again. What might be the problem and...

24 August 2016 9,913 3 View

Problem with RNA isolation from Breast tissue ?

I am trying to isolate RNA from Breast tumor and adjacent normal tissues (stored in RNAlater at -20 C) using Trizol method. After adding isopropanol , I am getting a transparent droplet like...

25 March 2016 2,797 4 View

Why do I get peaks in NTC in qPCR ?

I am performing real time pcr for ACTB. I am getting a ct value of 17-20 in my samples. However , i am getting a peak in NTC too with a ct value around 33-35. Also the dissociation curve for NTC...

01 November 2015 3,506 6 View

How can I select a good endogenous control for miRNA qPCR?

I have to perform qPCR for miRNA isolated from colorectal tumor and adjacent normal tissue samples. Is U6 snRNA a good endogenous control for qPCR?

16 June 2015 6,610 7 View

Do you know how I can process TCGA data?

I have downloaded the miRNA seq data for colon adenocarcinoma from TCGA. If I would like to see the expression of a particular miRNA across all the samples, how should we proceed?

07 April 2015 2,709 1 View

How should I calculate the amount of primers to be added from a primer mix?

I have 100 uM stock of forward+reverse primer mix. I have made a 10uM working concentration for the use in PCR. How much volume of this mix should i add to a 50 ul reaction volume so that i have...

02 April 2015 2,298 15 View

What is the appropriate blank for measuring RNA concentration on Nanodrop?

I usually elute RNA in TE Buffer but most of the protocols describe using water as blank and for the dilution of the samples. So, what is the appropriate blank to be used for RNA samples in TE buffer?

31 March 2015 600 6 View

How can I prepare nuclease free water?

I want to prepare solutions with nuclease free water to run RNA samples(isolated from Mirvana isolation kit) on denaturing acrylamide gel. It is given in protocol not to use DEPC treated TBE. So...

30 March 2015 4,807 10 View