What is the best housekeeping gene for miRNA analysis in qRT-PCR?

More Houtan Vafaeifard's questions See All

Can we use the microRNAs involved in an abiotic stress as a basis for identification of differential expression of microRNAs under a biotic stress?

For example, can we use the microRNAs involved in specific stress in a plant submitted on mirbase.org as a source for analysis of those microRNAs in other stresses? Thank you in advance for your...

30 November 2020 5,344 3 View

How do I increase the yield of my PCR product?

Hello, We would like to increase the yield of our PCR product. We are running a series of PCR reactions that is targeting ~1.1kb sequence. We begin each reaction with ~400pg of template DNA...

02 March 2021 4,029 3 View

Sample dilution for quantitative analysis using ELISA?

If the detection range is in ng/ml but the reference range is in ug/ml for a molecule or protein in serum or plasma .how to dilute and what is the initial volume to be taken for quantitative analysis

02 March 2021 7,670 3 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

How to address this issue in the analysis of Real-Time PCR results?

To dear Researchers, I was analyzing a series of concentration for estimation of Real-Time PCR efficiency. The concentration was 1:10. I used MS-excel to evaluate Slope. The result of slope was -8...

01 March 2021 8,683 4 View

No title

Does anyone have the experience of using Taq Man probes in the QIAGEN Rotar- Gene qPCR machine?

01 March 2021 5,311 1 View

Analysis of CRD two factor using R studio ?

Hello everyone. I have conducted the research on post harvest longevity of tomato crop using plastic packaging and different doses of chemicals as two different factors. The design of experiment I...

28 February 2021 8,189 3 View

Anybody with an idea on how do design specific primers for detecting tomato resistance genes tm-1, TM-2 and Tm2^2?

I am trying to identify these 3 genes among some tomato cultivar collections and after aligning some sequences from NCBI, I couldn't find unique sequences to target for specific primers. There...

28 February 2021 606 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

Runs of nucleotides in Sanger sequencing?

I performed site directed mutagenesis, transformation, and then I sent out plasmids for Sanger sequencing and found out that there is extension of DNA just before the stop codon. I am not sure...

27 February 2021 547 3 View

Aissa Saidi

Among several stable internal reference genes (doi:10.1371/journal.pone.0051502), you can use the COX gene PCR set as described in the papers below:

Article Development of a One-Step Real-Time PCR Assay for Diagnosis ...

Article The development of a validated real-time (TaqMan) PCR for de...

Dear Fahd Alhamdan, thank you for your feedback allowed me to open a window on microRNA applications in plant biotechnology. As Houtan Vafaeifard wants to use qRT-PCR in plants (Tomato: Solanaceae), I only suggested the COX gene (that I have already employed in qRT-PCR for the diagnostic of many plants diseases (ToLCNDV, Pepino, ToBRFV...), the suggested papers are still recommended by the American Phytopathological Society (APS). :)