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Questions related to Real-Time PCR
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 2,066 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 4,074 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 300 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 9,559 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 6,408 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 5,395 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 4,926 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 9,995 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 2,396 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 5,430 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 7,879 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 3,477 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 4,676 3 View
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 438 3 View
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 2,078 1 View
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 7,322 1 View
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 8,389 7 View
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 934 4 View
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 8,500 2 View
06 October 2019 1,022 6 View
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 299 4 View
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 1,478 3 View
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 5,422 4 View
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 1,772 4 View