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I will do a PCR for cloning, for this reason the primers have the sequence of the ERs at their ends, for this reason the Tm are high. It is possible to do a PCR with these high Tm primers and what...

03 April 2019 1,240 5 View

What's the best human macrophage cell line for estimation of the diferential gene expression of S. aureus during a infection celular model?

I need to compare the gene expression of S. aureus during infection celular model.

06 July 2018 1,897 2 View

I need a dye for the quantification by flow cytometry-based assay or cholorimetric method-based, can somebodyhelp me please?

I need to make the growth bacterial curve

09 October 2017 8,402 5 View

Do you know any protocol for the isolation of phages for staphylococcus aureus from nasal swap?

05 June 2017 6,378 1 View

When phage DNA phages are purified, how do we know if this is the phage DNA and not the host bacteria?

31 December 2015 7,358 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

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

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

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

Could not amplify CD28 and CD3z ectodomain from cDNA of Jurkat Cells ?

For my experiments, I have to express ectodomains of human CD28 and CD3z. As a source, I extracted RNA from Jurkat cells and verified the quality of RNA on the gel. I, then, prepared cDNA using...

23 February 2021 7,051 4 View

Why does the sum of reaction forces not match the force component of a Free Body Cut in Abaqus?

I am using Abaqus to simulate a plate with in-plane loading. The boundary conditions allow the right edge to move up and down, the displacements on the left edge are fixed. There is no rotation...

23 February 2021 711 3 View

What effect does the large temperature difference between the two primers have on PCR, including real time PCR?

What effect does the large temperature difference between the two primers have on PCR, including real time PCR? If I did not get a result from real time PCR, is it possible that my problems are...

21 February 2021 4,094 7 View