The primers that I have are amplifying lower concentration of DNA but not higher. Is this even possible? What can be done to fix this?

I am trying to do qPCR using some primers. The primers are specific for lower concentrations. However, I do not see any amplifications at higher concentrations. What could be the reason for...

06 July 2019 4,201 1 View

How much concentration of primers is ideal to use for sensitivity?

The DNA that is in my sample is less to be detected as compared to host DNA. So, I have to increase my primers concentration from 10 micromolar to more. So, was wondering how much concentration...

01 January 1970 4,725 4 View

Why are most greenhouse studies having small population and sample sizes as compared to the ones described in statistical books?

I am a beginner in experimental design so maybe I am not understanding the basics. In a class that I took , for a given population and confidence level and interval, the recommended sample size is...

01 January 1970 936 0 View

What is the usual concentration of Diaporthe fungus using DNeasy mini kit protocol?

Hello Everyone, I am using DNeasy mini protocol to extract DNA from at least 7 days old culture in PDA. When I tried it, several times I am getting less than 10 ng/micro liter of DNA. It is...

01 January 1970 1,393 2 View

I am trying to extract DNA of fungus from plant tissue and amplify the fungal DNA. Any suggestions on the protocol?

Can anyone explain how it works. The fungus lives in the vascular bundles.

01 January 1970 7,572 2 View

The housekeeping gene stability analysis?

I am doing real time PCR , relative quantification for a fungus that infects soybean. What housekeeping genes do I use for this? Are these for the fungus itself? I did not find them for my fungus...

01 January 1970 938 1 View

Can I prepare standard curve from DNA extracted from pure culture of my fungus for absolute quantification?

I am working on a fungus and was wondering if I can extract DNA from the pure culture of the fungus and measure the concentration of DNA and do a serial dilution for absolute...

01 January 1970 6,554 4 View

Can we analyse the reference gene expression without software mentioned in literature like Normgne, REST etc?

I am trying to calculate the reference gene expression stability for my qPCR assay since nobody has worked in case of my soybean. I am working on relative quantification of a fungus inside soybean...

01 January 1970 562 0 View

Trying to replicate the research paper , but the thermal cycling conditions of the primers do not work. Why?

I am trying to replicate using a set of primers mentioned in a paper Quantification of Fusarium solani f. sp. glycines isolates i... The thermal cycling conditions mentioned are 95 degree...

01 January 1970 1,488 9 View

What is your perception on using visual analysis or quantitative approaches to evaluate data from single-case research designs (n of 1 trials)?

Results of single-case research designs (i.e., n-of-1 trials) are often evaluated by visually inspecting the time-series graph and computing quantitative indices. A question our research team is...

03 March 2021 687 1 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

Research on Supply Chain Management in Indian context?

Hi, I am planning to apply for the PhD degree in the Supply Chain Mgt. with specific area of "Cold Storage warehouses" during Pandemics and wars. Where lock downs and shut downs are frequent....

02 March 2021 285 2 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 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

Dapi fluorescence protocol for colon cell lines ?

28 February 2021 2,534 2 View