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

Any help regarding small nucleolar RNAs (SnoRNAs) ?

08 September 2015 9,087 1 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

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

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

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

Are exogenous molecules transferred between human cells during doubling ?

Hello RG community, I wonderer whether exogenous molecules (e.g. reporter plasmids, miRNAs or other artificial molecules) are transferred between cultured human cells during their doubling ? So,...

21 February 2021 6,633 3 View

Has anyone encountered any problems with abm good microRNA primers for quantification of mature miRNAs?

I've been trying to quantitate mature microRNAs in 293T cells with and without 'X'. 2ug of total RNA (isolated by Trizol method) is converted to cDNA using All-in one microRNA cDNA synthesis kit...

17 February 2021 8,986 2 View

What is the best way to fix plasma derived miRNA for subsequent cDNA synthesis?

I need to quantify blood plasma derived miRNA through q-RT-PCR. For this I need to fix RNA content in my cDNA reaction. What is the best way to do this? Currently, I take readings of my plasma...

03 February 2021 9,330 3 View

What is the general miRNA yield one gets from the Qiagen miRNEASY serum/plasma advanced kit?

I am using the Qiagen miRNEASY serum/plasma advanced kit, using 400ul of frozen plasma. What is the yield you get on average? I am using nanodrop currently for quantification. I am getting...

02 February 2021 8,806 2 View

Which miRNA arm is a right target for analysis, 3p or 5p?

I'm working on selecting mouse miRNAs to analyze and stumbling upon one confusing issue, some of the papers do not specify which arm they analyzed and they just state something like " miR-34a was...

16 January 2021 3,792 4 View

Is negative controls necessary for western blotting?

I am trying to confirm my miRNA target by western blotting. I have a ds mimic and a ss inhibitor of my miRNA. Is it necessary to use negative controls for mimic and inhibitor? If yes, how can I...

09 January 2021 2,320 3 View

Can I predict miRNA by knowing the mRNA target?

Hello! I am trying to predict in silico the miRNA by knowing the target. Most of published research starts by knowing the miRNA and predicts the target using online tools. Are you familiar with...

31 December 2020 7,433 5 View

What are the possible drawbacks of using miRNA sponge to enhance the specificity of gene expression?

I am searching for miRNA repeats put between WPRE and 3'LTR of my lentivirus, in order to enhance the specificity of transgene expression in human neuron cells. But if clinically translation of...

26 December 2020 9,921 2 View