Any literature on cholesterol content of plasma membrane in various human cells?

Dear all, Could you suggest any review/paper for a comprehensive list of cholesterol content of plasma membrane in various human cells ? For example, difference in membrane cholesterol content of...

08 September 2018 1,009 2 View

Robust cAMP assay kit for Gi coupled GPCRs ?

Dear all, Can anyone suggest me a robust cAMP assay kit for Gi coupled GPCRs? I would like to use this kit for cell lines stably expressing GPCRs that are coupled to Gi (inhibitory G proteins)...

09 October 2017 265 3 View

How to get rid of colony in negative control after blunt end digestion followed by ligation?

I have digested my vector and PCR product with restriction enzymes A and B (both blunt end cutters). For ligation I have either the vector or vector + insert. Now as it is a blunt end digestion,...

02 March 2016 8,850 4 View

Direct protein estimation from cultures cells in a 96 well plate?

01 February 2016 7,864 7 View

what is the basic advantage of using Ni NTA agarose over Ni NTA superflow for membrane protein purification?

Dear All, I want to know what is the basic advantage of using Ni NTA agarose over Ni NTA superflow for membrane protein purification? I know the capacity of the two are different, but is that the...

31 December 2015 8,796 4 View

How do I normalize my MTT assay for reduced cell number in treated sample if my drug inhibits cell division?

Hello All, I am doing a MTT assay where because of drug treatment , the no. of cells in treatment is reducing to half of control. Now, after MTT assay how do i know the reduction in absorbance is...

09 October 2015 1,758 12 View

Why does 0.32M sucrose prevent clathrin-mediated endocytosis?

why does 0.32M sucrose prevent clathrin-mediated endocytosis?

05 June 2015 9,519 0 View

Bioinfo tool for determination of metabolic pathways?

Is there any bioinformatics tool to determine any metabolic pathway given the components?

01 February 2014 5,927 4 View

How to prevent inner glass plate breaking during SDS PAGE in PROTEAN II xi cell?

I am using BIO-RAD PROTEAN II xi cell for protein gel electrophoresis. I am using 15mA constant current for first two hours then 35mA constant current for the rest of the run. My inner glass plate...

03 April 2013 5,393 28 View

How does 67% Acetic acid in 33mM MgCl2 dissolves ribosomal proteins but precipitates rRNA?

Ribosomal proteins from ribosome is extracted best by 67% acetic acid in presence of 33mM MgCl2. What is the mechanism and how does acetic acid helps here? For reference : 1....

03 April 2013 6,184 1 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