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Questions related to Quantitative RT-PCR
I did prepare 1litre of comet assay lysis solution by adding sodium lauryl sarcosinate. For the assay used only 100 or 150ml and kept the remaining at 4 degree celsius for future use. After 2...
14 February 2023 2,433 4 View
As per the method described by Aebi (1984), extinction coefficient for catalase is 39.4 L/micromoles/cm. but in many research articles, the given extinction coefficient is 39.4 L/mM/cm and 40...
12 February 2023 1,360 1 View
Any optimized protocol for gene knockdown in THP1 cells using siRNA?
09 February 2023 6,658 0 View
I performed an immunohistochemical study and I have estimated the percentage of expression of different tumor markers, in this case, can I estimate fold changes with respect to the control ?
08 February 2023 3,757 6 View
Our main problem is that organoids embedded in agarose cannot be traced. Thank you!
29 January 2023 2,151 1 View
Hello I wish good health to all my friends. I had two questions. 1: Is free vitamin D in serum the same as vitamin D in VDBP in terms of immunological properties and can it be identified with a...
21 January 2023 4,093 1 View
Molecular identification of number of bacteria in saliva by using RT-PCR This technique gave us the only Cq but we need the starting bacterial level in saliva
18 January 2023 7,818 1 View
In analyzing data from my qPCR experiments, analysis on GraphPad is done using the ddCt, in presenting the analysis in graph, which is the most appropriate, the relative fold change or the ddCt?
06 January 2023 9,151 0 View
I used a KOD DNA polymerase for PCR that works very well. Then I used this enzyme to run a TaqMan probe real-time PCR but the results waere unexpected. I attached the results. The fluorescence...
31 December 2022 6,620 4 View
With good pictures?
27 December 2022 3,841 3 View
analysis of lipids in GC/MS
26 December 2022 279 1 View
Our oligonucleotides anneald stick-to-stick tail during PCR. Why another possible variants were not presented (or presented in very small amount)?
05 December 2022 6,650 1 View
Hi all, I am looking to test relative gene expression for my gene of interest in comparison to a reference gene. I realised, belatedly, I'd need a standard curve for every plate and I am wondering...
03 December 2022 1,543 0 View
Hi all, my name is David Previously I cloned a protein and everything was perfect. However now I want to clone this same protein but with a PelB sequence in its N-terminal for that I designed the...
02 December 2022 5,686 1 View
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 2,427 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 4,003 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 9,993 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 4,578 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 6,034 1 View
thanks
01 November 2022 9,215 2 View
Human dermal fibroblast passage easily or human foreskin ones?
30 October 2022 1,467 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 1,760 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 840 5 View
Is it fine to just store the sample at -80 C after treat it using detergent?
03 October 2022 5,155 1 View