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Questions related to Real-Time PCR
Hi dear fellows, I would like to perform ChIP-PCR on the promoter sequences of genes, but I am not quite sure how to design primers for the promoter sequences. Can someone please help me? My...
21 September 2020 7,518 6 View
We have tetanus toxin heavy chain fragment C (TTHc). It is suppose to have the ganglioside binding site for uptake. We can provide fluorescent TTHc. I am looking for someone who can show...
20 September 2020 7,130 0 View
We know that passive reference dye/ROX can help reduce variations of fluorescent values in samples. When utilizing the Rotor Gene-Q for the real-time PCR experiment, usually, the SYBR Green...
30 August 2020 10,197 5 View
Dear all, Recently we tried to compare mRNA expression levels of a gene via real-time PCR in 40 colorectal cancer clinical isolates and 10 normal mucosa samples. Interestingly, a reasonable...
28 August 2020 9,176 1 View
I would like to compare expression of my gene of interest (GOI) in non-treated leaves, stems, roots of my plant. In that case I don't have any control sample here and it's not possible to have...
10 August 2020 4,180 3 View
For comparing the quality of two products, I used the same sample and primers to do qPCR, the results are as follows. As the figure shows, Tm values are a little different. I wonder if this...
24 July 2020 7,442 3 View
Hello. I would be thankful if anyone could answer my question. We are Analyzing the expression of a certain gene in our research. We have extracted all the mRNAs in the cell and now I was...
23 July 2020 896 4 View
Why are CT values in the Realtime PCR hidden after the 35th cycle ? Why are CT values in the Real-time PCR hidden after the 35th cycle ? Where I noticed that the CT value had been positive and...
14 July 2020 3,781 9 View
Dears, I need help with the interpretation of why not obtain results in real-time PCR when I use Taq man SNP genotyping assay ID: rs 22756913 (IL17A) from Thermo fisher scientific where the...
24 June 2020 8,276 4 View
There are various methods of determining amplification efficiency as summarized under number 1 and 2 and in the bullets below: 1. From calibration curve slope as determined by: · Fit-point...
20 April 2020 9,151 3 View
Dear all, I would like to highlight a selected region of the synteny (collinearity) which only contains ~10 gene pairs. I have the gff file and the homolog pairs (links). What is the most...
15 March 2020 5,637 2 View
Hello Everyone, I wanted to run RT-PCR using onestep real-time PCR systems and SYBR green (ThermoFischer) to check the gene expression in bacteria. I wanted to directly use 50 ng of DNA samples...
11 March 2020 2,088 3 View
Hi I will be happy for your help. I'm doing real-time PCR with two primers called MT-TL1 and 18S, for mtDNA and houskeeping gene, and I got positive control on both NCT. I tried also different...
08 March 2020 4,097 4 View
Is it necessary to perform RT-PCR analysis of genes that show an increase or decrease? Or is it just for control purposes in RNA-Seq Analysis?
20 February 2020 321 5 View
I've been tried to align some histological slices on Amira but the software request a Stacked Slices file format. The main problem that I don't possess the z-values of the slices. I saw a...
18 February 2020 9,567 3 View
When we want to use Lanthanide-based DNA dyes (Eu, Tb)for time-resolved real-time PCR, the colors are limit. So how to develop more multi-color Lanthanide-based DNA dyes?
17 December 2019 6,429 1 View
I tried to scale down the component in reaction (from 20 µL to 10 µL using Bio-Rad Universal SYBR Green) for saving material. I wonder if I can get precise results or not with a down-scaled...
13 December 2019 5,412 3 View
We are intending to purchase a Real-Time PCR. Which instrument performs better? Whether to purchase Peltier based model or rotor based model?
08 December 2019 4,943 4 View
I have extracted total RNA from Lactobacilli and treated the samples with DNase I. When checking the -RT on qPCR, there was always gDNA contamination (appeared at cycle 33 or 35, automatic...
06 December 2019 10,009 3 View
I have some pictures of TEM from my samples and I would be grateful if somebody can help me to interpret them.
29 November 2019 2,406 6 View
I performed real-time PCR using an Applied biosystems machine. The double delta Ct values ( according to the machine) for the gene of interest after certain treatments were given as -5.12454;...
24 November 2019 5,453 3 View
Can we use money detector with UV lamp to detect fluoresceins such as FAM or FITC? For example product from Real-Time PCR with FAM or FITC probe?
20 November 2019 7,892 3 View
For example we have a bacterium containing two plasmids, one of which carries a target gene. Is it possible to calculate the number of copies of this particular plasmid performing real-time PCR...
18 November 2019 3,487 3 View
I am performing a qPCR standardization of some MGB probes. I have tried just a few times but I don't want to waste reagents, every time I tried the assay, there is a jagging line. I am thinking...
14 November 2019 4,696 3 View