Polymerase Chain Reaction

In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short... | Contact experts in Polymerase Chain Reaction to get answers

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Questions related to Polymerase Chain Reaction

(qPCR troubleshooting) Are these qPCR results reliable?

Hi, I conducted qPCR using probe included FAM and BHQ-1. At two annealing temperatures(60 and 65 C), the Ct value, R^2 and PCR efficiency in the amplification plot were also within the normal...

06 October 2022 6,728 2 View

Does a falciparum trophozoite have periodicity???

I observed in our clinic the trophozoite of falciparum malaria either late or early stage detected from 7 pm to 11 pm while early ring stage or merozoites dots with early ring lines are detected...

02 October 2022 8,645 0 View

Can doxycycline be used to treat inflammation?

if yes then what type of inflammation and how ?

01 October 2022 8,840 1 View

Is there any database for functional information of Enzymes ?

I am.performing LCMS analysis of my target sample (crude) for its proteomic analysis and After LC-MS analysis, i would need to check for properties and function of target proteins or enzymes...

24 September 2022 7,048 4 View

How to prepare strawberry samples for FRAP assay? i mean what is the procedure of sample preparation like homogenization or other technique?

before the starting of FRAP assay it needs to be preparation like homogenization or fractionations. my question is exactly focused on the homogenization methods. if there is any other technique,...

22 September 2022 8,413 0 View

Reamplification of Gel Purified PCR Product gives same non specific bands. How to get rid of non-specific band?

I did an overlap-extension PCR and got the following gel image (purple image + neb 1k ladder). The target band size is 1.9kb, which was the brightest band. Afterwrds, I cut the 1.9kb bands and...

09 September 2022 8,393 0 View

What would outliers indicate on a Ramachandran plot?

I am doing a paper on a glucoamylase structure and have generated a Ramachandran plot. The plot has no outliers and 93.27% of the amino acids are in the favored region. I understand that the amino...

08 September 2022 6,280 1 View

Which is the mechanism of S. aureus USA300 being resistant against kanamycin ?

dear colleagues , i have got a quite simple question on today , which is the mechanism of S. aureus USA300 resistance against kanamycin at very high concentration 64 ug/mL ? could it be...

03 September 2022 7,930 0 View

How to Analyze Antimicrobial resistance Genes and Bacteria in real surface water? What things should I need? can you please explain in detail?

I am looking to analyze ARB and ARG from real surface water of marshland.

30 August 2022 453 3 View

Why green fluorescence appearing in EtBr agarose gel while loading my plasmid construct containing viral protein.?

We have checked buffers, Etbr and running buffer.....we are wondering why we are getting this green fluorescence in well....

26 August 2022 4,722 0 View

Can I call two SNPs in 'perfect linkage disequilibrium' with D'=0.9995 and r=0.99?

I analyzed two SNPs for linkage disequilibrium using SNPStats. It gave D'=0.9995 and r=0.99 but when I analyze my SNP data Manually, always these two SNPs are either present or absent together in...

22 August 2022 4,401 3 View

¿In an alignment between two nucleotide sequences of two different strains of a same organism with 82% of identity we could talk that are same gene?

I just have do an multiple sequence alignment (about 15000 sequences) between different strains of a bacteria for determine its conserved regions, but near of 1% of sequences have 80 or 85% of...

12 August 2022 3,622 2 View

How can an association be made among genes by a Transcription Factor that binds to several of those genes?

If a Transcription Factor is discovered to bind to a few genes, is it possible that those genes have a relation between them? Is there a possibility that just because there is TF binding to 2...

11 August 2022 3,715 2 View

How to use CRISPOR software for design guide RNA?

I want to know tutorial help to design and how to choose suitable g for knok out

30 July 2022 4,043 2 View

Detecting Phytophthotra spp.?

Hi! can you advise me on some method to find Phytophthora spp in soils? I tried the baits technique (following the advices of https://bsppjournals.onlinelibrary.wiley.com/doi/10.1111/ppa.13312),...

30 July 2022 5,813 7 View

In logistic Regression, what does it mean when your P-value for a variable is not significant but your Odd- Ratio CI cross 1?

I am working on a logistic regression model, with a binary outcome and my model look like this: Intercept: coefficients is -2.034 Variable: coefficients are 0.031, and the P-value is 0.130. Not...

27 July 2022 6,042 7 View

Inexplicable consistency in cq value, what could the problem be?

Good day, I have a question related to particular results with qPCR. The performed assay has been fully validated but is now encountering some unexplained results. It's about the inconsistent...

26 July 2022 1,276 0 View

How to find biologic effect of specific mutations in Dengue genomes (i.e. aminoacid level or antigenic properties once folded)?

Hi all! I have found specific mutations in dengue genome sequences apparently related to the infection outcome. Here my questions: How can I find if this mutation is synonymous or...

25 July 2022 3,108 2 View

Why do the smaller DNA markers narrow at the bottom of the gel?

This is a 1% agarose gel in 1xTBE with 1xTBE running buffer, visualized on UV transilluminator using ethidium bromide. The markers are 1KB+, and the rest of the signals are my plasmid miniprep...

22 July 2022 7,491 0 View

Could specimen preserved in bad condition get polluted by other sample in the same container?

I got some lizard specimen which had been stored in 95% alcohol for over 15 years at room temperature. Also, the lizards were all being preserved in the same container, while their legs were...

21 July 2022 622 2 View

Can I use HOMER to perform a Motif Discovery analysis with GWAS (SNPs) results?

I have a list of significant SNPs and would like to perform a Motif Discovery and Transcription Factor analysis.

14 July 2022 8,440 4 View

Why it is problematic to obtain PCR amplification using the product of overlap extension PCR as template and the same terminal primers?

I have been setting up overlap extension PCR to join four fragments of size 500, 2140, 1300 and 500 base pairs. I am adding equal amount of DNA in molar ratios and using Taq Polymerase with traces...

13 July 2022 4,190 0 View

Different combinations of housekeeping genes give me different results? What is the best amount of HK genes in qPCR?

I was trying different housekeeping genes in qPCR analysis and every combination of them gave me another result. When I choose one HK, two, or three, the results were completely different. I am...

13 July 2022 2,726 2 View

Did anybody have a solution for "dirty" sequences?

Hi all, I'm working with analyzing DNA taken from prey. My goal is to find a predator who killed. I'm isolating DNA from swabs (saliva), with GeneALL micro DNA kit, then I'm making PCR with D-loop...

07 July 2022 2,363 4 View

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