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Questions related to Quantitative RT-PCR
Hi I'm doing my master thesis and having a hard time optimizing my PCR reaction. I'm trying to add 2 concentrations of MgCl to my mastermix. I already have 1,5 mM MgCl in my master mix but need it...
24 November 2022 2,310 1 View
Avian embryos (while still in the shell) were exposed to hepatocarcinogens. Hepatocyte isolation by collagenase. In culture, over time, these “monsters” appear. Although by now the focus of my...
24 November 2022 3,875 1 View
For your information, I am using ABI StepOne Plus machine. The housekeeping gene is Beta-actin. However, the results showed its melting curve and fluorescence plot look good. Appreciate it if...
14 November 2022 9,840 1 View
I have values of two treatment groups with similar gene target and all 3 groups have the same beta actin value. Treated 1 = 21.00 cq and Treated 2 = 20.50 cq, while both treatments having the same...
10 November 2022 4,454 3 View
I'm struggling with understanding and troubleshooting qPCR issues with cycles 1-3. The rate of change for the cycles 1-3 is so high, it is tricking the software into thinking that is where the...
02 November 2022 5,890 1 View
thanks
01 November 2022 9,133 2 View
Human dermal fibroblast passage easily or human foreskin ones?
30 October 2022 1,355 4 View
Hi everyone! I would like you to show off your most successful strategies to acquire bacterial / archaeal high quality intact RNA from environmental samples / mixed community (liquid...
20 October 2022 1,644 0 View
The first few qPCR runs the primers worked and I was getting signals in my samples/positive controls. However, now when I run another qPCR with the same primers I get no signal. I've tried using a...
18 October 2022 719 5 View
Is it fine to just store the sample at -80 C after treat it using detergent?
03 October 2022 5,007 1 View
It is not compatible with MS as buffer in the mobile phase for LC, but is it OK to have in sample? The concentration is around 70mM. Thanks.
07 September 2022 6,419 0 View
I performed a titration treatment on cells in 2 replicates and then detected an antibody in the supernatant using ELISA. I have my corrected measures and standards, sample concentrations and SD...
27 August 2022 4,254 3 View
I am looking at copy number variation from frozen skin samples. The gene of interest sometimes has two bands of positive droplets across different samples. What causes this?
14 August 2022 8,421 0 View
I am trying to plot Weibull distribution using Origin. In the y-axis, I am getting percentages. In most of the publications, the Y-range is different starting from some -ve value to positive (-2...
30 May 2022 4,442 1 View
I am currently using an ABCAM AGP competitive ELISA kit with a dilution factor for the samples of 40,000 which was suggested by another lab. The problem is that the blank has a lower OD than the...
11 May 2022 5,863 0 View
Are there any consortium that can ship a cDNA clone in Europe?
04 May 2022 4,316 2 View
Good day, could anybody suggest what could be the reason of a additional big band need upper marker? If we exclude contamination. I am using Agilent sure select XT protocol for targeted DNA seq of...
05 April 2022 6,965 0 View
We currently have a study done with RNA-seq analysis. But the raw counts in our data show a large difference, up to 10 fold higher in the raw counts versus the lowest one. But after normalization,...
01 April 2022 3,047 5 View
I ran an assay with Cy5 probe using Bio-Rad CFX96. Why are some of the curves less intense than some of the other curves? The smaller curves are using cDNA and the larger curves are using plasmid...
01 April 2022 4,626 3 View
I am doing photodegradation experiments on Contaminants of Emerging Concern, and surprisingly I see an increase in TOC from the beginning to the end of the experiment! How this could be possible?...
22 February 2022 6,771 11 View
When preparing the reaction mixture for cDNA synthesis, I messed up while adding the template. So, I had to downscale the total reaction volume for cDNA synthesis to 10 uL. I use the Thermo...
17 February 2022 1,083 2 View
I was tring to check the differential binding/association of my protein to genes using mutant S. pombe. For this, I used a Flag tagged normal strain and a Flag tagged mutant strain. I did...
12 January 2022 7,234 0 View
I performed WB with 15 ug/10 ul protein sample for 1 lane My expected band is indicated in red marker in the image. But something wrong with my band, always. Band is connected by strong dots...
25 September 2021 8,538 0 View
I am studying photocatalysis of Polystyrene nanoplastics (22 nm) with TiO2 18 nm (%80 Anatase, %20 Rutile) under UVC in DI water (15 mL solution in 20 mL vials. Concentrations: 20 ppm PS, 20 ppm...
18 September 2021 7,211 5 View