How to improve RNA extraction using Trizol?

I have been extracting RNA using trizol method from plant tissues but sometimes during the extraction procedure there is low pellet formation by which the quality and quantity of RNA gets...

09 October 2018 9,101 0 View

How to increase the intensity of bands in sqRT Pcr analysis for gene expression?

I have tried increasing the no. of cycles as well as checked the annealing temp but still i get a faint bands wherein i m suppose to get bright ones. even it appears same in case of housekeeping...

09 October 2018 8,475 2 View

What might be the reason that during quality using 0.8% gel electrophoresis i got a single thick band but not two separate bands as its seen normaly?

I extracted RNA from the fresh tissues using RNA kit form GCC Biotech, normally during the Quality check using electrophoresis it showed Double band but today in my case i could get only one...

08 September 2018 3,595 4 View

Can someone suggest me what might be the possible reasons for the appearance of smear, faint and no bands in semiquantitative RT-PCR gene expression?

I use the primers synthesized by molbiogen and all the pcr components purchased freshly and cDNA synthesized shows expression in housekeeping gene. I tried in normal DNA i got faint bands for the...

07 August 2018 7,707 2 View

Does housekeeeping gene show two different band size when the cDNA is prepared using reverse primer instead of Random Hexamerase?

I extracted the RNA and prepared cDNA using the reverse primer of OsActin instead of Random hexamerase and did a PCR for the same, but to my surprise i found two bands of different size of two as...

07 August 2018 462 5 View

Is it possible to do acid digestion of plant material in big borosil testube using water bath optimizable to 100 degree celsius?

Acid digestion of rice roots in 70% conc. HNO3.

05 June 2018 757 2 View

Please Can anyone suggest me a method of powder sample preparation from rice root tissue for XRD analysis of presence of elements?

I wish to study the presence of elements in rice roots under heavymetal toxicity.

03 April 2018 1,673 2 View

Reasons for the use of CaCl2 solution in preparing Aluminium stress solution after growing in nutrient solution. why do we use CaCl2 solution?

Reasons for the use of CaCl2 solution in preparing Aluminium stress solution after growing in nutrient solution. why do we use CaCl2 solution?

01 February 2018 3,764 0 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