What's the optimal temp. for hot start Taq polymerase? and do we need an special PCR condotion or not?

I've ran a PCR using Hot start Taq polymerase instead of normal taq polymerase by mistake, but as the result i have found some pure and non-sharp bands, may any one help me to find why it has...

11 December 2019 4,912 3 View

What's the effective protocol for DNA extraction from Filamentous fungi?

I have tried some protocols in order to extracting DNA from Filamentus fungi, such as proteinase K, RNxplus, TRizol, 95 Degrees of c. heat and ....but still its a failure. how should I extract...

06 July 2016 5,620 9 View

In the case of circulating miRNAs expression measurement, which one is more credible? the Serum or Plasma?

03 April 2016 4,143 4 View

Housekeping Genes: which one is the best?

there are some genes accepted as internal control for gene expression analyses but nowadays some researchers say expression of these housekeeping genes are varying from a tissue to other or their...

05 June 2015 5,485 8 View

What are the sings of necrosis?

is it ok to say necrosis will increase the activation and expression of anti-apoptotic genes and pathways? I have some problem in my project which lead me to think that necrosis will increase the...

03 April 2015 4,161 4 View

How can I find the related lncRNAs of a gene?

I'm looking for regulators of a specific gene, want to find sncRNA ( like microRNAs) regulators and LncRNA regulators of the gene... about the microRNAs I've find some useful info using miRWalk...

02 March 2015 8,876 4 View

Has anyone extracted DNA from cultured apoptotic cells using TRIzol kit?

I had extracted total RNA of cells according to the manufactures and tried to extract DNA of the same sample respectively, but i see no bound on electrophoresis gel. I followed all steps one by...

02 March 2015 2,836 5 View

Should I centrifuge the cells after thawing them in order to remove the DMSO from the medium?

All we know, centrifuging right after thawing the cells is harmful. But in the other hand, remaining the DMSO in the medium can hit the cells, too. So shall I remove the DMSO or it's better to...

11 December 2014 1,311 23 View

How much FBS do I need to add to the medium in order to prevent cell division in 96-well plates?

My MCF-7 cells are dividing in 5% medium, but I need to keep them alive and prevent them from division...

08 September 2014 1,457 4 View

How to calculate the IC50 for my MTT assay?

there is many formulas for calculating the IC50 value but I can not use them because I don't know what does mean x, x', Y and Y'. there is a formula Y = a * X + b, IC50 = (0.5 - b)/a. but I don't...

07 August 2014 4,404 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

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