Clostridium spp. primer qPCR sequence?

31 December 2019 6,662 3 View

I change the annealing temperature, should I change the melt temperature in qPCR?

31 December 2019 8,200 3 View

QPCR melt curve problem with gut tissue DNA?

I am analyzing an intestinal microbiota of rats by qpcr. I had already done the efficiency of my primers and analyzed samples of DNA extracted from feces and everything went well. Good melt curve...

31 December 2019 593 3 View

Same CT for diferent dilutions/concentrations?

I am working with DNA extracted from feces. I made a qPCR using concentrations of 100, 75, 50 and 20ng final. The CT value was very similar between dilutions between 29 and 30. My positive control...

10 November 2019 7,803 8 View

Can polysaccharide contamination in DNA extraction samples cause the DNA band to appear as degraded on agarose gel (electrofluorese) reading?

The picture of gel is there

04 May 2019 184 4 View

No title

I already used a lysis buffer, but it did not help.

03 April 2019 5,560 3 View

Best protocol for extraction of bacterial DNA from feces with CTAB method?

I am trying to extract bacterial DNA from feces and cecum from mice, to then do PCR. I initially tried with phenol-chloroform and did not succeed. I had better results with CTAB, but the DNA band...

01 January 1970 6,572 3 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

How to increase a model accuracy ?

dear community, my model is based feature extraction from non stationary signals using discrete Wavelet Transform and then using statistical features then machine learning classifiers in order to...

03 March 2021 6,994 5 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

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 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

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