How much 100mM of Gluconic acid can change pH of a buffer?

11 December 2014 3,260 1 View

How can one decide on DNA amount, plating amount and screening size in mutant library construction?

08 September 2014 5,798 4 View

What are the most suitable conditions for E. coli DE3 sonication?

08 September 2014 246 4 View

What is the reason for my huge solvent peak?

08 September 2014 2,483 19 View

What is the most important feature/parameter for considering a gene as a pseudogene?

If its reading frame has a start codon and no stop codons in between, is there any chance that it will make a protein?

07 August 2012 9,325 6 View

What is the trick to working with low volumes in a french press?

I am using constant systems French press. There is lot of volume loss and frothing happening. I am working with bacterial cell suspension and volumes are around 1-2 ml. All kind of suggestions are...

07 August 2012 9,524 5 View

Rule out the possibility of tRNA/rRNA in smallRNA population?

I have some smallRNA candidates. How can I confirm that they are tRNA/rRNA, any online software/program/tool available?

05 June 2012 5,923 2 View

Is there any tool for analyzing intergenic regions?

In order to filter out sequences on the basis of size, and stop codons.

05 June 2012 6,731 0 View

Using western blot apparatus for northern blotting?

My lab has a regular set up for protein electrophoresis and blotting. I want to use the same for RNA blotting. Which recipe is the best and least time consuming to make my whole setup RNase free?...

05 June 2012 2,155 4 View

What methods are there other than northern blotting for small RNA validation in prokaryotes?

I have some intergenic small RNA candidates in prokaryotes which I want to validate.

05 June 2012 2,571 7 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

Why is the efficiency of viral mRNA electroporation in PBMC low?

Hello, I have been struggling with the electroporation of PBMC with HIV-I mRNA. Although the quality of the RNA generated is good, and the handling of the cells in meticulous, the frequency of...

27 February 2021 9,233 3 View

What is the best concentration of cDNA to be used for qPCR?

I have synthesised cDNA from 500ng/ul of RNA. The concentration of the cDNA Is about 5000ng/ul. Should I dilute my cDNA or just use it as it is for my qPCR? I usually use 2ul of cDNA for a 20ul...

24 February 2021 8,618 3 View

Recommendations on homogenizers for RNA isolation?

I will be performing RNA isolation with frozen lung biopsy samples and am in the market for a tissue homogenizer. I am wanting a homogenizer that will fit in a 1.5ml tube. I was looking at the...

24 February 2021 3,623 3 View

RNA-seq DEGs downstream analysis and reporting in literature?

I used EdgeR to get the list of DEGs (differentially expressed genes) from my liver RNA-seq experiment. I have done the gene overrepresentation and GSEA based pathway analysis. I am currently...

23 February 2021 8,969 1 View

Is it efficient to do liquid biopsy to investigate the gene expressions of the tumor?

To do liquid biopsy and look for specific gene expressions there are so many published papers. Thera are different protocols and procedures used to isolate circulating cell free RNA or mRNA from...

23 February 2021 7,326 5 View

RNA degraded after good nanodrop readings?

23 February 2021 8,850 2 View

Is there a way to extract total RNA and protein from cell culture at the same time, without denaturing protein?

Hi all, Is there a way to extract RNA and protein from the same cell culture sample? Because we need to use the protein for gelatin zymography at later time, we are looking for a method that can...

22 February 2021 5,341 5 View