Real-Time PCR

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Questions related to Real-Time PCR

How to get the SNP flanking sequence(for Prunus mume, Japanese apricot)?

I already get SNP candidates for our target traits, then I would like use Real-time PCR based genotyping platform to check F1 population. For establishing the marker, the SNP flanking sequences...

10 November 2019 468 3 View

Old TEM microscopes - anyone recognizes this model?

Dear TEM&RG community, does anyone recognise the old TEM on the image, probably from end of the '60s? The microscope was installed in Slovenia, also wall-text suggest that. Based on our...

07 November 2019 2,099 1 View

Is there any significant limitation in doing real time pcr over pp65 antegenemia aasay??

A baby with congenital cmv infection tested positive using realtime pcr with 1000+ copies/ml of DNA. Further doctors ask for po65 assay why?? How do they go for treatment??

18 October 2019 7,344 1 View

I need a trick to remove DNA contamination from extracted RNA samples?

Hi, everyone I am extracting a total RNA from rat brain samples for real-time PCR studies, but unfortunately, there is DNA contamination in gel electrophoresis. I tried to remove it and I failed....

10 October 2019 8,404 7 View

Gene expression in a cross-sectional study?

Dear colleagues I have gathered human adipose tissue to assess the association of leptin gene expression with dietary intake of fat. I have measured leptin and GAPDH(housekeeping) gene expression...

10 October 2019 944 4 View

Can I use gen A's exon for a positive control to detect a deletion of gen B's exon via Real-time PCR?

Dear everyone, I'm working on a project to detect the deletion of exons. We are working with Realtime PCR and using Livak's method. But we don't have a positive control of the target gene, we only...

10 October 2019 8,511 2 View

I need a trick to remove DNA contamination from extracted RNA samples?

Hi, everyone I am extracting a total RNA from rat brain samples for real-time PCR studies, but unfortunately, there is DNA contamination in gel electrophoresis. I tried to remove it and I failed....

06 October 2019 1,061 6 View

Having linear amplification curve with SYBER Green qPCR?

I am profiling miRNAs using SYBR Green real-time PCR. Some of the miRNAs show only a linear amplification curve without any exponential part. In these cases, the amplification starts at very early...

02 October 2019 315 4 View

How can i extract miRNa from saliva samples efficiently ?

Recently we decide to evaluate the miRNA expression levels in saliva sample as diagnostic biomarker of cancer . We used TRIzol extraction methods but the CT of real-time PCR for U6 gene was...

09 September 2019 1,513 3 View

Real-time PCR statistical analysis. What to do when homogeneity of variance fails?

Ddear colleagues I'm performing a statistical analysis of real-time-PCR data by the delta delta ct method. Basically I'm analysing the expression profile of a target gene in non-inoculated...

09 September 2019 5,443 4 View

Can anyone guide me about AriaMx Real-time PCR System and it's results?

Dear all, Actually, I am working with AriaMx Real-time PCR System. I read the protocol of this device and following that to work but unfortunately, I don't know why I can not get any...

20 August 2019 1,781 4 View

I extracted DNAs from vegetable samples stored at -80C and got very little DNAs. Is my store condition of samples is wrong?

Hi. I'm testing the existence of parasites on the surface of commercial vegetables by using real-time PCR method. I extracted genomic DNA of vegetable samples using QIAamp stool mini kit. Before...

08 August 2019 1,360 3 View

Can I compare between dCt of the samples in Real-time PCR? As there is no control in the experiment to calculate ddCt?

the experiment is about comparing between different plant fungi treatments on the gene expression of the plant.

08 August 2019 6,046 7 View

Any suggestion for low cost Real-Time PCR?

I am planning to buy a real time PCR instrument for my laboratory in Chennai, India. Budget is my primary concern and I am in a great dilemma to decide which machine might be the best choice for...

08 August 2019 8,126 3 View

How do I get consistent CT values from two different PCR machines?

I have some DNA controls I would to use for real-time PCR but I'm sure at what concentration to use these controls. I have the following volumes 1. Bordetella pertussis (19000 copies/ul)...

08 August 2019 2,600 5 View

What is the reason for the variation in reading of standards set from a stock of a plasmid by the Real-Time PCR?

Dear All, To measure the DNA copy number of HBV virus, I prepared a standard series from the HBV-containing plasmids stock (10 +5, 10 +4, 10 +3, 10 +2, 10 +1), and after reading with Real-Time...

08 August 2019 2,951 4 View

Is the oncogene transcription level stable between different passages in hUC MSC ?

Hi guys, I found the mRNA level of oncogene （for example: C-MYC，C-FOS...) is unstable between different passgaes in hUC-MSC using realtime PCR. Is this a Q-PCR technical issue or is these genes...

24 July 2019 4,316 3 View

What can be the possible reason for the signal observed in NTC during TaqMan probe based qPCR (Real Time PCR) assay?

Hello everyone, I am carrying out TaqMan probe based Real-Time PCR assay. I am able to observe signal peak for the NTC sample. However upon agarose electrophoresis I was unable to see any band...

18 July 2019 6,714 6 View

Does anyone have experience with using the Illumina ECO real-time PCR system for HRM analysis?

Does the Illumina Eco Real-time PCR system require calibration for HRM (SNP detection)? The system manual mentions 'no temperature shift or calibration required'. However, I need to be certain...

07 July 2019 9,809 1 View

Is the oncogene transcription level stable between different passages in hUC MSC ?

07 July 2019 9,628 3 View

Hello to evreyone i have a question about RNA isolation from plasma with Trizol. how we should do it to reach out a good 260/280 ratio?

I did it According to the following protocol but the 260/280 ratio was 1.3 and 260/230 was 0.22 I know that it is not good for doing real-time PCR: 1)400 Microliter plasma + 1000 microlitr...

07 July 2019 2,849 9 View

Can anyone help me with determining protein expression using quantitative Real time PCR?

I am working on a sperm specific protein, I want to determine its concentration in control and infertile. Can it be done using real-time PCR and how?

25 June 2019 9,405 9 View

Can anyone help me with determining protein expression using quantitative Real time PCR?

I am working on a sperm specific protein, I want to determine its concentration in control and infertile. Can it be done using real-time PCR and how?

06 June 2019 779 10 View

How to transfect (stable) pancreatic cancer cell lines (Panc-1, BxPC-3, AsPC-1, MIAPaCa-2) with plasmid DNA?

Hello everyone, I am currently trying to stably transfect the pancreatic cancer cell lines Panc-1, BxPC-3, AsPC-1 and MIAPaCa-2 with plasmid DNA having a G418 resistence gene. I tried FuGene HD...

06 June 2019 4,674 5 View

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