EMSA troubleshooting. Protein-DNA complex not migrating, cold probe lane with multiple bands. Can anyone help?

I'm running an EMSA to show the DNA binding activity of recombinant proteins versus the Wild type. Previous assays run by my research group using a different test system show the protein-DNA...

24 February 2021 8,325 1 View

Physical-chemical formation of stalactites/stalagmites. what promotes the formation/growth of these structures?

I know, or think I do, the basic idea behind these natural structures, as from what I know they form due to water dripping from the top leaving a salt deposit behind. But started to wonder about...

22 February 2021 1,400 4 View

Why is my qPCR suddently not working?

I've been working with qPCR for some months now. I amplify 2 genes (PSEN2 and HN1), and the protocols are optimized for both and have been working fine. The other day I amplified HN1 for some...

21 February 2021 3,437 3 View

Can the intensity of this broad band in XRD mask other crystaline phases present?

I obtained a XRD diffractogram with significant broad band at low 2theta levels, also around 30 2theta another smaller broad band appears. It all points out to the existence of an amorphous phase....

19 February 2021 8,414 4 View

Can ethanol be used as an analgesic?

If ethanol has analgesic properties, why isn't it used for clinical purposes?

18 February 2021 8,227 4 View

Acid phenol:chloroform (25:24:1) for RNA extraction instead of (125:24:1)?

Hi everyone! I am about to extract RNA for RNA sequencing assays and my Invitrogen kit/protocol...

14 February 2021 1,755 3 View

How to measure a Neural Network complexity?

Does anyone know how to measure the complexity of an artificial neural network? Could you provide me references? Thanks a lot!

25 January 2021 2,710 10 View

Are there any public datasets of consumer complaints about service industries at the European level?

06 January 2021 1,718 2 View

What is the best method for removing power-line interference (PLI)?

In the paper A wavelet-based method for power-line interference removal i... we propose a new method for removing PLI. This method consists in identifying the ECG and noise frequency range for...

29 December 2020 9,233 3 View

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16 December 2020 9,657 5 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

MiRNA qpcr problem?

Dear All, mirna primer showing some problem in the melting curve? any idea why? As attached is the melting curve. The forward sequence is obtained from miRBase and reverse primer is universal.

28 February 2021 5,008 4 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

Can MicroRNA cross the blood brain barrier?

In diseased conditions, many genes and gene products are visible in the bloodstream. So my question is whether we can find traces of microRNA which is involved in disease and disorder in blood serum?

24 February 2021 7,064 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