Why does it use TE as negative control in qPCR multiplex?

Dear all, I have a question. I do not understand why does it use TE buffer as negative control in qPCR multiplex? Is it the same if I use the mix as negative control? Thanks so much Giorgia

06 July 2018 6,969 2 View

Protocol PCR primers narG1950m & narG2050m?

Did someone used these primers in PCR or qPCR? Because I have a problem to perfom the PCR and I obtained the no appropriate band in the samples and NTC. Thanks Giorgia

31 December 2017 8,664 3 View

Which method do I use to find the best gene reference to use un the RT-qPCR?

Hi, I need to find a gene reference to use in the gene expression of cellulolytic genes in Ascomycete. Which gene reference is possible ti use? Or which is the method to find it? Thanks Giorgia

05 June 2017 3,518 1 View

Did not see amplification in ITS qPCR?

02 March 2017 6,751 4 View

RT-qPCR protocol one or two steps?

Is best to prepare the RT-qPCR in one-step or two-step?

10 November 2016 5,669 9 View

Is BEST analysis (PRIMER 7 software) a multivariate analysis?

I try to analysis the best correlation between the biotic and environmental dataset. My question is: if I have one variable for the biota dataset and more variable for the environmental dataset,...

01 February 2016 1,753 2 View

In PRIMER, is it possible to downweight certain taxa in multivariate community datasets to reflect uncertainty around identification??

I have a dataset with about 80 different species. As usual, some species are very easy to identify with certainty whereas others are more difficult, which means that I am less certain of my...

03 March 2021 8,066 4 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

PACYC PCR not working after several attempts. Would anyone have any suggestions?

So, I have been trying to run a pACYC PCR which will be used later on for a Gibson Assembly. However the PCR is not working. I have already tried gradient PCR and changing extension time; however...

02 March 2021 1,146 2 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 calculate the annealing temperature of a primer pair?

I have to amplify a gene and my primers just reached. The Tm for Forward primer is 64.2, and that of reverse primer is 65.5. Can some one suggest how to get the best annealing temperature? Thanks...

01 March 2021 360 7 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

Isolates of Hymenoscyphus fraxineus?

Dear Dr. Cai, my name is Simone Prospero and I work in the team of Phytopathology at the Swiss Federal Institute for Forest Snow and Landscape Research (WSL;...

01 March 2021 1,133 1 View

How to remove the residual PDMS on the silicon mold?

Incompletely cross-linked pdms are difficult to remove.

01 March 2021 798 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