How can i get a positive control for my PCR test on Alfalfa mosaic virus ?

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Why Non specific bands of PCR reaction appear while using specific primers for tobamoviruses?

i have done RNA extraction and cDNA for tobamovirus then i made a PCR reaction using primer pair Tob uni1/Tob uni2 according to the protocol in Letschert et al., 2002 ( annealing temp. 60C for 45s...

11 December 2019 7,406 3 View

Why my specific primer match to give cDNA but it does not work in PCR?

I work in plant virus and I use a specific primer to make the cDNA (I do not use oligo dT or hexamer) and it works giving me a sharp band (photo in attachment) But when I made the PCR using these...

07 August 2019 1,504 3 View

Why ELISA reading decrease with time!!! ??

I made ELISA test and read it after 50 min , the results made sense and it was OK But i reread it after 1h and 45 min but the results are less than the first time . Why this happened?

06 July 2018 5,904 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

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

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

01 March 2021 5,311 1 View

How vaccines of covid 19 work?

Millions ask how vaccines of covid 19 be successful and safe, its here to explain generally how vaccine of pfizer work ,mRNA vaccines teach our cells how to make a protein that will trigger an...

28 February 2021 2,723 4 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

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

Can thermal cycler machines be problematic during PCR?

Hi, my question is about the heating of thermal cycler machines and I hope some of you had experienced a similar thing previously. There are two thermal cycler machines in the lab(BioRad) and for...

26 February 2021 4,777 4 View

Calculating volume of DNA required from DNA concentration?

Hi I am a bit confused. They are asking me to find out the volume of DNA required in ul (a total of 30-100 ng for genomic DNA) from the DNA concentration in the nanodrop reading which was 404.8...

26 February 2021 5,029 2 View

Thugiang Thi Bui

I have one suggestion, you can contact the DSMZ in Germany, they have a huge collection for virus samples. Their website is: www.dsmz.de

I think they can help you to find a solution.

Have a nice day,

Govinda Sharma

If you have a reference sequence, you could have it synthesized. For example, IDT does longer dsDNA synthesis at really reasonable costs. Since it's a standard for a PCR test, you probably won't be synthesizing anything too long so it will probably be fairly cheap.