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 7,904 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 5,845 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 2,113 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 8,430 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 2,630 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 1,071 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 3,314 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 7,301 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 9,330 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 6,437 7 View

Is there a problem with my RNA pellet?

Hello, I am currently having problems with RNA extraction. I am using mouse liver (C57BL6J), and I have extracted RNA from mouse liver before. Before this experiment, my final RNA pellets were...

11 August 2024 7,082 3 View

Strugglling with m6A dot blot any suugesstion ?

I have been doing the m6A dot blot for a while with no improvement, I am extracting the RNA, and I can see the dots although the three biological replicas give a different reading on the memberan...

10 August 2024 8,539 5 View

RNA Extraction Using Hot Borate Method No Longer Working?

I've been performing RNA extraction on cotton petiole tissue for a few months now using the method described in the following paper, a derivative of the typical hot borate method...

08 August 2024 9,882 2 View

Does Anyone have expertise in in vitro transcription and RNA pull down assay?

I am currently working on LncRNA; to know the lncRNA-protein interactions I want to do RNA pull down assay, so I need to design primers with T7 promoter. I need assistance in this regard.

07 August 2024 6,622 1 View

E.coli contamination in human RNA seq data ?

Recently, we observed that 99% of the sequences in our RNA-seq data corresponded to the E. coli genome. Despite multiple DNAse treatments after RNA extraction and ribosomal depletion, we were...

06 August 2024 807 3 View

How to assess the osmotic power of a substance?

Hello, I suspect that a molecule I am using is causing bacteria to experience osmotic shock. What chemical properties should I look for to compare this capability with those of other substances?...

06 August 2024 977 1 View

RNA later for the preservation of RNA in fecal samples at room temperature for one day (37°C)?

I am planning to collect human fecal samples for metatranscriptomic analysis using MGI. These samples are from indigenous people living in a region with high temperatures. I will have access to a...

06 August 2024 1,367 3 View

Do you have good tips for seaweed tissue preservation in the field for post RNA extraction?

I will be with my students collecting seaweed samples in a marine farm and later we will process this tissue for RNA isolation and further sequencing. Does anyone have tips on how to collect the...

04 August 2024 501 2 View

What is the acceptable p-value cutoff for GO enrichment analysis ?

I have an RNA-seq data that I have analysed using Limma-voom and have extracted the gene IDs, log2FC and the p-values. At p value < 0.05, I have over 10,000 DEGs, however, when I run the GO...

31 July 2024 225 2 View

Inquiry on Maximum Nucleic Acid Volume for 2.5 mL Liposome Solution?

I am currently working on a project involving liposomes and need to determine the maximum volume of siRNA that can be added to a 2.5 mL liposome solution with a total lipid concentration of 10...

30 July 2024 6,420 1 View