Abnormal notched curve in qPCR ?

Dear all, Any comment on the reasons of appearance of notched qPCR curves ? See attached. The machine in Roch LC480

09 October 2018 1,922 0 View

Abnormal notched curve in qPCR ?

Could You help explaning the reasons why a notched curve after qPCR appears ? The machine is Roch

09 October 2018 5,951 0 View

Which tool to be used for building miRNA-mRNA interaction ?

I have a list of miRNAs and a list of predicted target mRNAs. The mRNA targets were created by other ways. I am searching for a tool to test whether an interaction (binding) could exist between...

08 September 2018 6,148 4 View

Speculation about virus titers in a disinfection study for un-measured contact time ?

Dear All, Could any body help me in finding the best statistical/model to see the un-measured titres of virus in a time points that extend beyond the ones I measured. For instance I am testing...

07 August 2018 3,735 2 View

In gene bank submission, what is the reason of "your seq. contains stop codons" ?

I am trying to submit sequences of COX-1 gene(mitochondrial gene) to gene bank. My sequences is almost 100% identical to a published one. It return a message saying that they contain stop codons...

04 May 2018 984 6 View

Which type of regression to be used ?

What is the suitable regression to be used if I have binary independent variables and also the dependent variables ar binary ? i.e. yes/no ? Details: I am checking whether the presence/absence of...

09 October 2017 634 6 View

Why do I loos some DE miRNAs upon combining some time points together during the DE analysis?

I have a time series experiment and want to compare DE miRNAs between these time points and the Control. I realized that I loos some DE miRNAs upon combining some time points together and conduct...

08 September 2015 1,466 1 View

How can one determine miRNAs in ducks and Turkey ?

04 May 2015 946 2 View

Can I use U6 primer assay from Qiagen as standard gene in miRNA detection in chicken samples ? Both are 100 % conserved ?

04 May 2015 3,231 1 View

How to make in-situe detection of miRNA in cell line?

10 November 2014 9,131 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

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

How does the injection volume impact LLOQ and ULOQ in regression analysis (LC-MSMS)?

Hello, I have run into an issue that has sparkled a debate at work: is the LLOQ/ULOQ impacted by injection volume under the following circumstances: 1) Same dilution factor of all samples 2)...

23 February 2021 9,159 3 View

RNA degraded after good nanodrop readings?

23 February 2021 8,850 2 View