Contact experts in Quantitative RT-PCR to get answers
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Questions related to Quantitative RT-PCR
Hello, My current lab is analyzing qPCR data in a method that is frankly, very incorrect. Can anyone recommend a step-by-step example for analysis of significant differences between treatments,...
01 March 2021 2,216 0 View
I have question as described in the title - how can I add residues to protein, with the PyMol Builder, and mark them as non-organized? PyMol gives options to add them as beta-sheets or...
28 February 2021 5,608 2 View
I ran RT-qPCR for a control and a treated group and it appears that my gene of interest was not expressed at all in any of the subjects in the control group. So I was wondering if I could...
21 February 2021 6,488 10 View
I've been reading up on how to best represent qPCR data, specially on answers given on this site by Dr. Jochen Wilhelm. I understand box plots are the way to go, but I still don't understand them...
12 February 2021 7,561 3 View
Hi everyone, I'm looking for some help troubleshooting a recent qPCR issue I've encountered. I am using fluorescent Taqman probes and a qPCR master mix that includes ROX as a passive reference....
09 February 2021 559 3 View
We use the NEBNext Library Quant Kit for Illumina to determine our Illumina library concentrations. Before taking the library to qPCR, we usually run the library on Bioanalyzer to get an idea of...
28 January 2021 7,118 4 View
Hello.....I am reconstructing the amplification curves from raw fluorescence data out of the .xml file exported from the LC480II cycler. I cannot understand how to apply color compensation off a...
27 January 2021 876 0 View
We're trying to isolate primary intestinal epithelial cells from 13 days old Balb/c mice. Although our protocol allows us to efficiently isolate them with minor fibroblast contamination, when we...
22 January 2021 5,162 0 View
I am looking for isolation (FACS) and cell culture protocols for rat Type I alveolar epithelial cells. I have read "Isolation and Culture of Alveolar Epithelial Type I and Type II Cells from Rat...
10 January 2021 850 0 View
I am trying to determine what best time-point to harvest my cells, HMVEC, after transduction with long non-coding RNA to determine changes in mRNA levels, specifically VEGFR2/KDR. I am capable of...
20 December 2020 4,994 2 View
Hi, I have been doing Reverse Transcriptase qPCR on some liver mRNA samples. I am targeting liver inflammation genes like Col1a1, alpha SMA, IL6, IL1B and etc... Even though mRNA concentrations...
18 December 2020 6,366 3 View
Hello everyone, I have fabricated a stretchable antenna that uses silver nanowire (AgNW) as a patch and Polydimethylsiloxane (PDMS) as a substrate, but I encountered a problem when installing SMA...
06 December 2020 8,383 4 View
I wonder if the positive control primers and probes authorized by the US CDC for amplifying human RNAse P are specific to the RNAse P chromosomal DNA. My thought process is that if primers are...
02 December 2020 2,532 2 View
I have seen COVID19 test LoD ranging from 180 to 600000 NAAT Detectable Units/mL So a rapid antigen test with LoD of TCID50 1.13 x10^2 per mL and RT PCR TCID50 1x10^-2 per mL (600 NDU/mL) and a...
29 November 2020 1,406 3 View
Hello everyone. I have collected intestinal biopsies from stem cell transplanted patients in RNAlater and wish to quantify neutrophils only via qPCR. As we know that the mucosa of gut is filled...
26 November 2020 2,875 3 View
So the lab that I work in generates a lot of AAVs and we titer them via QPCR. However, I've been noticing that the entire time, the efficiency has always been very low (anywhere from 40-80%) with...
23 November 2020 4,731 3 View
Hello everyone, My experiment is to assess the expression levels in cultured cells after the clock has been reset, or in mouse liver by qPCR. I have multiple timepoints and have performed...
22 November 2020 979 2 View
I used triplicate for a sample in qPCR, and the Ct values are 26.0, 26.02 and 31.1 It is obvious that 31 is an outlier which may be resulted by technical error and I want to exclude it in the...
18 November 2020 4,574 3 View
Hi! I have to perform a qPCR experiment in which I would like to evaluate the expression of 13 genes (plus 3 HKG) on 10 samples. I must carry the runs on a 96-well qPCR plate since I need to...
15 November 2020 5,653 4 View
Hello, we are currently working on absolute quantification with SYBR Green qPCR. We excised from a plasmid containing the cDNA, a fragment, which will be amplified by the primers we used in our...
11 November 2020 969 4 View
I want to use molecular docking to find the interaction of amino acids and solute carrier family 7 member 3 (CAT3), but the PDB of CAT3 is not available in the protein data bank or any other...
02 November 2020 3,457 12 View
I am attempting to plan a qPCR experiment in which I evaluate the expression of multiple genes in various brain regions of fetuses, as a function of location in utero (intrauterine position,...
01 November 2020 5,916 1 View
I would like to see RNA-protein interaction following transcription process inside the cell. I am wondering if there is any protocol similar to protein co-immunoprecipitation,which can bind the...
28 October 2020 3,155 3 View
While reviewing my qPCR, I noticed that I had different melt peaks for samples run in triplicate. Is this something I should be worried about, or is this normal? I've uploaded a word doc showing...
27 October 2020 7,440 2 View