Increase RNA concentration?

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How to separate protein-RNA complex?

I am purifying protein which has RNA in its bound form. How can I remove this bound RNA from protein without affecting structure of protein. RNase treatment is not much effective.

31 December 2018 7,802 4 View

Protein complex in Native PAGE?

I am trying to separate protein complex and protein monomer in native PAGE but protein is not able to move inside the gel and it get stuck at well only. I tried with 5% acrylamide, 5% glycerol,...

04 May 2018 5,720 5 View

Any expereince about gluteraldehyde crosslinking?

I am trying to crosslink my protein by gluteraldehyde but after crosslinking multimeric form of protein is not able to run on SDS PAGE even after 6 hrs long run at 90V. Please share your experience.

02 March 2018 2,032 4 View

PSUMO PRO vector cloning?

Hello, Does anyone have experience in ligation of >1.5kb gene in pSUMO PRO vector or any other vector. Please give suggestions.

10 November 2017 8,341 4 View

Ligation independent cloning ?

Can anyone provide basic experimental setup for ligation independent cloning method. I am not able to understand the basic mechanism behind this method.

08 September 2017 2,534 14 View

Contamination in Rosetta?

I am inoculating Rosetta strain in LB broth but everytime culture got contaminated. Even after fumigation of lab. Please suggest any reason or solution.

03 April 2017 992 11 View

Restriction sites known but getting unknown gel band. How?

I digested my circular plasmid having 1 RE site for BamH1 and 1 for Bsa1. So I supposed to get 2 bands but getting one more low molecular weight band. Please explain any reason.

11 December 2016 3,227 16 View

Rosetta cells in autoinduction media?

Hello I have cultured rosetta cells in ZYP5052 autoinduction media. It is showing very slow growth. Does anyone have some experience with this media? please share .

11 December 2016 7,177 3 View

How addition of Arg + Glu increase protein solubility?

If Arg and Glu are added to purified protein it increases the solubility of it. Why?

08 September 2016 9,249 4 View

Solubility of Protein on lower temperature?

I have read in experimental procedure that lowering the temperature will translate more soluble protein. What is the reason behind it ?

07 August 2016 6,348 7 View

Does any one know how i can prepare liver tissue with transmission electron microscope with good fixation?

i have problem in preparation of liver tissue with bad cell organelles and how i can adjust osmolarity

03 March 2021 8,890 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

Calculating volume of DNA required from DNA concentration?

Hi I am a bit confused. They are asking me to find out the volume of DNA required in ul (a total of 30-100 ng for genomic DNA) from the DNA concentration in the nanodrop reading which was 404.8...

26 February 2021 5,029 2 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