RNA extraction and gel electrophoresis ?

Dear all, I am trying to isolate RNA from bacteria for RNA seq. I have used fresh samples for extraction as well as 3 days stored (-80) samples. After gel electrophoresis I didnt observed 3...

11 December 2018 704 11 View

Selective marker (antibiotic resistance gene)

I am trying to knockout gene in bacterial strain (genome sequence is available) so I must choose the first selective marker but I found that this strain is resistant to aminoglycoside, Blactum...

09 October 2012 1,776 4 View

Quantitative analysis of cellulose in Medium ?

Can anyone suggest me good method for the quantitative determination of cellulose in Broth and medium after fermentation other than DNSA test?

08 September 2012 3,330 4 View

Enzyme Immobilization in polyacrilamide beads

Immobilization of penicillin acylase in porous beads of polyacrylamide gel (...

06 July 2012 2,917 7 View

Separation of nanoparticles according to their shape?

I want to separate nanoparticles according to their shape. I have read that electrophoresis and density gradient centrifugation give good results. Could anyone recommend what the best method for...

06 July 2012 7,879 13 View

Can my bacterial strain synthesize PHA?

I want to check whether or not my isolated bacterial strain is able to synthesize PHA (Polyhydroxyalkanoates). How can we identify PHA content in the bacterial cell by GC chromatography? What is...

05 June 2012 7,720 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