Real-Time PCR

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Questions related to Real-Time PCR

There are difference between 25uL and 50uL of final volume in qPCR?

I’ve been performing, so far, my qPCRs reactions in 50 uL of final volume; however, at doing in this way, I have spent a lot of material. In this sense, I would like to know if I could perform my...

03 March 2018 9,179 4 View

Who could provide primers for quantification of retinoid X receptor alpha (RXRA) mRNA?

Hello RG community, I want to determine the effect of miRNAs on the 3´UTR of retinoid X receptor alpha mRNA and therefore want to analyze its mRNA level in HepG2 cells. I want to go and check for...

03 March 2018 1,100 5 View

How to check tissue specific distribution of mRNA using Real-time PCR?

I just want to check the expression of a particular mRNA throughout the different tissues (Liver, Kidney, Brain, Muscle, Gills, Intestine, Ovary, Skin, Spleen etc.) in a control/untreated fish. If...

28 February 2018 2,315 6 View

Housekeeping genes for ER stress in cancer cells?

We are trying to quantify different genes involved in ER stress and autophagy in glioma cells by semiquantitative real-time PCR. The cells are treated with tunicamycin or thapsigargin or are...

27 February 2018 750 2 View

Real-time PCR instruments ?

We are going to purchase qPCR instrument to our new genomic platform; Which the better choice regarding specifications, medium throughput applications and price between this instruments : qPCR...

21 February 2018 2,914 3 View

RT-PCR for p38 MAPK pathway genes in breast cancer cell lines?

I have some active compounds and I want to check the effect of those compounds on p38 MAPK pathway genes (p38, JNK and ERK1/2) or may by AKT/PKB and PTEN in MCF7 and MDA-MB-231 breast cancer cell...

07 February 2018 6,220 3 View

Real-time PCR instruments ?

We are going to purchase qPCR instrument to our new genomic platform; Which the better choice regarding specifications, medium throughput applications and price between this instruments : qPCR...

02 February 2018 3,611 5 View

How to check tissue specific distribution of mRNA using Real-time PCR?

I just want to check the expression of a particular mRNA throughout the different tissues (Liver, Kidney, Brain, Muscle, Gills, Intestine, Ovary, Skin, Spleen etc.) in a control/untreated fish. If...

02 February 2018 5,367 7 View

Housekeeping genes for ER stress in cancer cells?

We are trying to quantify different genes involved in ER stress and autophagy in glioma cells by semiquantitative real-time PCR. The cells are treated with tunicamycin or thapsigargin or are...

02 February 2018 584 2 View

Are there simple and efficient methods for extraction both RNA and DNA from the same sample of Gram-positive bacteria?

I am working with highly mutagenized Bacilli species and wish to measure expression level of my favorite genes by real-time PCR. As I know, there are no reliable internal reference genes for...

14 January 2018 1,725 0 View

Is there a way to check the contamination of primers with cDNA?

I want to check the contamination of primers with cDNA, because in real time PCR their peaks are not very good and have extra peaks. Moreover, the negative control without cDNA (NTC) has a peak...

12 January 2018 4,440 10 View

Why do I get two or three peaks in melting curve of real time PCR?

Hi every body, I want to set up real time PCR for one gene, but after doing it I get two or three peaks in melting curve of real time PCR. The designed primers for my gene is very specific. After...

04 January 2018 1,456 13 View

Housekeeping gene normalization in real-time PCR is it always necessary?

Hi, i want to analyze the expression of SNAIL 1 and 2 in various cancer cell lines by real-time PCR. Is it always necessary to perform a housekeeping gene validation with geNorm according to...

01 January 2018 7,368 3 View

Are there simple and efficient methods for extraction both RNA and DNA from the same sample of Gram-positive bacteria?

I am working with highly mutagenized Bacilli species and wish to measure expression level of my favorite genes by real-time PCR. As I know, there are no reliable internal reference genes for...

01 January 2018 9,987 1 View

Does anyone have any articles on detection of SEN virus, using Real-time PCR Techniques?

01 January 2018 3,927 2 View

Antibiotic resistance genes in wastewater ?

i am work with antibiotic resistance genes in wastewater with Real Time PCR analysis, like Tetm gene, Sul1 gene, Intl1 gene. who can help in some questions ? thank you Ammar

01 January 2018 7,551 2 View

Does anyone know about MGB-EQ (Eclipse) probes?

I synthesized an existing probe in the literature (FAM-MGB). The probe is marked with the quencher MGBEQ (eclipse) (Company Eurofins Genomics). However, the probe did not amplify anything, it may...

12 December 2017 2,938 0 View

Luciferase activity increase expression but real-time no expression/decrease ?

Hi, has anyone exprience, the luciferase assay resulted in increasing the expression but by the analysis using real-time it is low/no expression at all. Is it because post-transcriptional...

11 November 2017 2,183 4 View

Any specific methods or tools for discriminating two antigens from two genotypes ?

Dear there is a plasmid which has two nucleotide sequences from two genotypes ( C2 and A types ) We have to prove each of sequences are expressed well not only one sequence As you know, the...

10 October 2017 2,885 4 View

Which lobe of the mouse liver should be collected for real-time PCR?

Hi everyone, I am a bit curious about the protocol of collecting the mouse liver for real-time PCR. As I would like to check the expression of the gene under two different conditions. However,...

10 October 2017 8,314 0 View

I am trying to operate a real-time PCR. How can I find a repetitive element on the genome of a specific organism?

09 September 2017 7,837 2 View

Possibility to get sequence information of primers premixed in a ready to use PCR kit/master mix?

Hello, I use a commercially available real-time PCR kit for poultry virus detection. I have used this kit for some preliminary experiments but need the primer sequences for further research. Is...

09 September 2017 5,196 3 View

Please What is the best protocol for making standard curve from bacterial genomic DNA using Real time PCR?

I want to compare the Traditional CFU count of bacteria with Real-time PCR method, but I m having some issues with the making standard curve. So please if anyone is willing to help I will really...

09 September 2017 3,924 3 View

Possibility to get sequence information of primers premixed in a ready to use PCR kit/master mix?

Hello, I use a commercially available real-time PCR kit for poultry virus detection. I have used this kit for some preliminary experiments but need the primer sequences for further research. Is...

04 September 2017 1,644 3 View

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