Restriction enzymes with genomic DNA?

Hello, Is there anyone has any experience with using restriction enzymes with DNA before doing PCR? in case having a pure extracted DNA

07 August 2018 9,264 4 View

Can using the same rapd primers for fungi and plant ?

07 August 2017 7,586 7 View

Can I detect the mtDNA in total genomic DNA without specific producer for mtDNA extraction?

09 October 2014 6,123 6 View

Can I detect the chloroplast DNA in total genomic DNA without specific producer for chloroplast DNA extraction?

09 October 2014 955 2 View

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08 September 2014 9,965 4 View

PCR Troubleshooting

07 August 2014 2,488 2 View

What is the annealing temperature suitable for the PCR reaction?

I have primers, but the problem is they have different Tm with a big range. The forward is 70 the reverse is 63.

04 May 2014 6,824 23 View

Can anyone explain this "plant contains genes that homologous in structure and function to genes of MT in animal and fungi"?

Does this mean they have the same genes? If they are homologous, can I use the same PCR primers?

03 April 2014 5,748 3 View

What types of RNA should I extract for reverse transcriptase PCR, miRNA or large RNA (Total RNA)?

What types of RNA should I extract for reverse transcriptase PCR, miRNA or large RNA (Total RNA)?

03 April 2014 3,187 8 View

Can I use the same PCR primers with different DNA organisms?

Sometimes when I am looking for some microorganisms primer genes in the web I found some of them, but have been used with different DNA organisms, can I use the same primers with different organisms?

02 March 2014 2,946 12 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

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Genome-wide gene identification protocol ?

Is there any book chapter/book, webpage or research article available to understand genome-wide gene identification?

28 February 2021 8,095 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

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 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