What are principles of Real Time PCR?

i need to know: Principles Protocol The best kits available with high efficiency results How to read and interpret the results?

31 December 2018 2,234 0 View

How can response to reviewer comment to remove a confused point or to prove a finding?

What the best way in Discussion section to remove a confusion on bacterial pathogen which has never recorded in the certain area and by PCR gave the same product size for a bacterium of our study...

04 May 2018 5,231 4 View

How can I manage large DNA sequences in DNA alignment?

During Blastin search, I found many sequences with Large sequence. (about 2105803 bp ). I faced a problem in adding them to DNA Alignment and in each time a message appeared "Warning: about to...

02 March 2018 2,076 7 View

Is there a site helps me to draw a map for use in epidemiological study to illustrate the geographical distribution of the disease?

I am working on epidemiological study, which aimed to study risk factors of infectious diseases distribution, spread and re-emerge in endemic areas. For this I am asking how can I draw a map...

02 March 2017 9,751 0 View

What are advantages studying the percentage of homology in protein sequences?

I have a revised manuscript sent for publication and the reviewer had asked from me insert the percentage of homology of protein sequences then discuss, however, I discussed the DNA sequences...

03 April 2016 3,575 6 View

Does anyone have experience with LAMP protocol instead of PCR assay?

I mean what are the principles of LAMP and How can I prepare my Lab for it ?

03 April 2016 3,546 2 View

What are the advantages of using amino-acid sequences in comparison to using nucleotide sequences for phylogenetic analysis?

I need know how can I use and interpret the amino acid sequences for phylogeny for the same homologous region

02 March 2016 7,205 16 View

I need a teaching video for Glanders and Strangles in equine describing epidemiology, clinical signs, diagnosis and control?

02 March 2015 5,882 3 View

Can I deposit the two different sequences of PPR virus regarding F and N genes in Genbank under the same accession number?

I have two sequences of PPR virus of two genes (F and N genes) (complete Cds).

09 October 2014 4,876 4 View

Does anyone have experience with the Kappa test?

08 September 2014 6,734 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

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 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

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 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

What is the ideal efficiency for a qPCR?

hello everyone, I need to do standard curves for my qPCR, what is the ideal efficiency range? I tried a primer (Mglu2 receptor) that gave an efficiency of 90.2%. Is it accepted?

28 February 2021 1,254 3 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

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

Has anyone used the Illumina ITS1 primer sets?

Hi everyone, Illumina provides a list of primers to amplify with high taxonomic coverage the ITS1 region for further fungal sequencing, but I cannot find the exact amount necessary of each...

25 February 2021 6,969 3 View

Why do the PCR bands from the cDNA templates (extraction from the cell after transfection) show different size from the band of the plasmid template?

After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...

25 February 2021 5,712 3 View