Contact experts in Real-Time PCR to get answers
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Questions related to Real-Time PCR
Hi all! I need some help, as I've been trying to troubleshoot and struggling with this for more than a few months. I have DOPC + 05% Texas-Red-DHPE liposomes that I need to fuse into planar...
15 December 2021 3,329 1 View
I tried staining 3D cancer spheroids using F-Actin stain but the F-Actin filament were not well distinguished as lines. It looked like mitochondrial or lysosomal stain. Any ideas why it turned...
06 October 2021 3,731 13 View
I have Ct values of my housekeeping gene, and i have ct values of 2 genes of interest. I want to show the level of these genes relative to housekeeping gene and relative to each other. I basically...
26 May 2021 9,465 4 View
To dear Researchers, I was analyzing a series of concentration for estimation of Real-Time PCR efficiency. The concentration was 1:10. I used MS-excel to evaluate Slope. The result of slope was -8...
02 March 2021 9,029 4 View
I Know that RT-PCR is Reverse Transcription PCR when the starting material is RNA. Going for reverse transcription first (RNA to cDNA) and then it could be conventional PCR with the need for gel...
12 February 2021 6,969 4 View
Hello everyone. I am about to search for the expression of genes including p53 in methylated and non methylated HPV samples. Should I extract DNA from the methylated samples and use SYBR green...
05 February 2021 2,864 3 View
Dear all, I wanted to run HRM ( High Resolution Melting) using Corbett Rotor-Gene 6000 Real Time PCR Machine. When I opened the HRM folder of software , there was two option for running including...
03 January 2021 3,941 6 View
Currently I process a commercial Multiplex Realtime PCR kit on Quantstudio 5 system, but almost 20% of the runs has a distorted amplification plot (see pictures attached below). Does anyone...
15 December 2020 7,976 3 View
High-Resolution Melting (HRM) analysis is a relatively new, post-PCR analysis method used to identify variations in nucleic acid sequences. The method is based on detecting small differences in...
27 November 2020 4,893 4 View
Hi all, I am trying to set up my standard curve for qPCR analysis, presence absence of D. septosporum, using a QuantStudio 5 real-time PCR system and associated software. I am using a 5X...
26 November 2020 834 3 View
I am working on quantstudio 5 system with diagnostic test kit - AZF Microdeletions REALTIME PCR genotyping kit (master mix is aliquoted to every separated pcr tube, I only added taq polymerase,...
20 November 2020 4,945 3 View
I have been using mfold web server for RNA secondary structure prediction, but it is no longer available since the 1st of November. I need a reliable and easy tool alternative to it or a...
03 November 2020 1,053 7 View
Hengzhi Shi , Minwei Li , Xiaocui Huang, Chaoqun Yao , Xueqiu Chen , Aifang Du , Yi Yang PMID: 32731054 DOI: 10.1016/j.vetpar.2020.109193.
31 October 2020 9,536 2 View
Hi, I want to multiplex some reaction with dual label probes. I check the probes on FAM, and HEX channel. When I change the dye for ROX channel I don't see any fluorescent. I ordered the probes...
20 October 2020 1,167 5 View
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 7,495 6 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 10,167 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 9,150 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 4,151 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 7,427 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 875 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 3,762 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 8,249 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 9,130 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 5,626 2 View