How to design primers for miRNA quantification by qPCR?

More Ram Babu Undi's questions See All

Is it possible to validate a miRNA target in a test tube by providing all the essential factors, and how can we determine the half life of a miRNA?

To validate a miRNA target mRNA can we do it in vitro by adding everything in a test tube? If not, what are the difficulties and how can we overcome this?

08 September 2014 981 4 View

Can anyone explain how to measure mitochondrial copy number using realtime PCR approach?

I am using realtime PCR approach to measure mtDNA copy number with mitochondrial tRNA or rRNA and nuclear B2M primers. I isolated DNA from both the samples control and treated then I did real...

06 July 2014 7,645 8 View

Can anyone explain why my MTT assay is giving off a brown colour instead of purple?

When I perform an MTT assay after adding DMSO, only a brown colour is appearing. I am working with suspension cells and I want to know the cytotoxicity of a compound. After 24 hours of seeding...

11 December 2013 7,328 15 View

Can anybody explain how to design primers for a mature miRNA ?

I want to check the miRNA levels and miRNA transcript is short in length. How to design primer for a specific mature miRNA?

11 December 2013 9,940 7 View

Can anybody give me protocol for mitochondria staining using mitotracker?

I want to stain the mitochondria in suspension cells by using mitotracker red (Invitrogen). Can anybody explain protocol to observe them in fluorescence microscope?

11 December 2013 829 12 View

Why is protein aggregating during concentration by amicon?

Buffer composition is 50mM tris pH7.5(p.I 6) 150mM Nacl (tried upto 300mM) 5mM BME 1mM pmsf 10% glycerol. Even it is also aggregating after NI -NTA purification but I am diluting protein then it...

11 December 2013 9,452 6 View

Can anyone please recommend a protocol for analysis of surface expression of a CD marker using flow cytometry?

When I am performing an experiment I could not observe the any signal. Usually I use FITC tagged antibody but I am suspecting that I have problem with sample preparation and cells may be damaging.

08 September 2013 8,117 14 View

Does anyone have CMK cell line?

I am working on the megakaryocyte development. I need this cell line for my study.

02 March 2013 5,341 1 View

Can anyone send me a detailed procedure of miRNA cloning in a mammalian expression vector?

11 December 2012 659 0 View

Anyone having idea about VN primer for miRNA primer design ?

How to design VN primer to attach with universal reverse primer

05 August 2024 2,116 3 View

Why did the authors extrapolate a phenotype that they experimentally proved in one bacterial strain across the whole genus of the organism?

I aim to be as skeptical as possible regarding whether a pair of orthologous genes results in the same phenotype in their different but related bacterial organisms under similar environmental...

05 August 2024 6,787 4 View

Why my colony PCR results of my recombinant bacterial not showing any results?

I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....

05 August 2024 2,570 3 View

What is the acceptable p-value cutoff for GO enrichment analysis ?

I have an RNA-seq data that I have analysed using Limma-voom and have extracted the gene IDs, log2FC and the p-values. At p value < 0.05, I have over 10,000 DEGs, however, when I run the GO...

31 July 2024 225 2 View

Inquiry on Maximum Nucleic Acid Volume for 2.5 mL Liposome Solution?

I am currently working on a project involving liposomes and need to determine the maximum volume of siRNA that can be added to a 2.5 mL liposome solution with a total lipid concentration of 10...

30 July 2024 6,420 1 View

Why cannot i find my protein on cell surface after antibiotic selection of expressing plasmid?

I cannot confirm cell surface protein expression by flow cytometry even after transfection and antibiotic selection of cells. Does it take long for proteins to express on the cell surface? the...

28 July 2024 3,178 3 View

Are these cassettes suitable for expressing PETase mutant in E. coli?

I created two potential gene expression cassettes (constitutive and inducible) for expression of a mutant PETase gene on PeptiCloud using the version tree feature, which allows users to create...

28 July 2024 7,559 1 View

Optimal siRNA Volume for Complexation with Liposomes in a 2.5 mL Solution with 10 mg/mL Lipid Concentration?

How much volume of siRNA can I add to 2.5 mL of liposomes with a total lipid concentration of 10 mg/mL?

27 July 2024 6,920 1 View

If my gene of interest has high GC content can it be problematic in sequencing? What kind of error is expected with GC rich gene sequences??

Gene sequencing related trouble shooting

25 July 2024 4,149 2 View

What is the function of fusion tag GAMGSGIQRPTST?

I found that several expression constructs of published protein structures containing an N-teiminal fusion tag GAMGSGIQRPTST. What's the founction of this fusion tag?

23 July 2024 1,353 4 View

Shibin Mohanan

Real-time PCR quantification of precursor and mature microRNA.

Methods. 2008 January; 44(1): 31–38.

doi: 10.1016/j.ymeth.2007.09.006

Alexander Sasha Vlassov

Applied Biosystems team will help - they have a plenty of "ready to go" TaqMan assays for miRNA (misc forms) plus can do custom design