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Questions related from Robert Rekawiecki
Have any of you performed MeDIP? After many problems in adopting Methylation Sensitive High Resolution Melting and Methylation Specific PCR methods in my research to determine DNA methylation in...
30 October 2024 2,168 0 View
I am performing methyl-specific PCR to determine the level of DNA methylation in the promoters of the isoforms A (PGRA) and B (PGR) of the bovine nuclear progesterone receptor gene. For each...
26 August 2024 1,351 0 View
Can anyone recommend polyclonal or monoclonal antibodies for Tet methylcytosine dioxygenase 2 (TET2) Tet methylcytosine dioxygenase 3 (TET3) DNA methyltransferase 3 beta (DNMT3B) Ideally, they...
28 June 2024 3,636 1 View
We would like to perform Methylation-Specific PCR using TaqMan primers and probes. With this method, we wanted to determine the DNA methylation of the promoters of the A (PGRA) and B (PGRB)...
29 February 2024 7,451 0 View
Has anyone used Promega's bioluminescent MTase-Glo Methyltransferase Assay kit on 96-well plates? I am looking for a protocol that calculates the amount of individual reagents for this type of...
10 January 2024 8,643 0 View
Does anyone use the MS-HRM method to determine methylation in a given locus of DNA? What software do you use to design primers for this method? The most recommended is Methyl Primer Express...
02 November 2023 4,111 0 View
I am looking for a precise method to determine the level of DNA methylation at a given locus. I am working on material from cows and I wanted to adopt the Methylation-Sensitive High-Resolution...
26 September 2023 4,721 1 View
Have any of you used MTase-Glo Promega kit for DNMT activity determinations? Do you need to purify DNMT as the protocol states or can you use isolated nuclear protein extract? Did you follow the...
13 July 2023 1,674 0 View
Can anyone recommend a company that can make or sell bovine fully methylated and unmethylated control DNA? Unfortunately, I have not found such controls commercially available for cattle. I would...
05 October 2022 1,672 2 View
Do any of the cells have genomic DNA that is 0% methylated? I found scientific reports indicating that peripheral blood mononuclear cells (PBMCs) may exhibit such properties. In my research, I use...
24 March 2022 8,226 5 View
Have any of you used Galaxy (https://usegalaxy.org/) to analyze NGS RNAseq sequencing data to search for new splicing forms of RNA? I am looking for a pipeline for data analysis in Galaxy. Which...
20 December 2021 4,053 1 View
I would like to isolate nuclear proteins from bovine endometrial tissue. The most recommended homogeniser for this kind of isolation is Dounce homogeniser. However, this homogeniser did not quite...
24 February 2020 3,450 0 View
Dear researchers I am looking for a procedure how to construct a vector for promoter deletion analysis. Especially, I mean how to generate deletion fragments of a gene promoter for further...
10 April 2019 7,402 8 View
I would like to design primers for HRM methylation analysis. Did any of you use the Methyl Primer Express software to design HRM primers. The software has two options for designing such primers...
04 April 2018 3,683 4 View
I would like to perform detection of apoptosis with using Annexin V (detecting translocated phosphatidylserine) or Mitotracker Deep Red (stains mitochondria in live cells) in luteal cells of a cow...
11 December 2017 8,167 3 View
Is there a possibility to collect cells after cell culture for further cytosol/nuclear fractionation. Cells were harvested in the middle of the night, and the procedure of cells fractionation...
30 October 2017 9,885 1 View
I performed real-time PCR on luteal cells from cows which were treated with various factors for a period of 6 hours. While for western blot, cells were stimulated with the same factors but for a...
01 January 1970 4,872 4 View
Dear friends. When I tested progesterone receptor coregulators during an oestrus cycle in a cow, I got some surprising results. The peak of protein expression occurs before the peak of mRNA...
01 January 1970 9,327 1 View
Is it always the case that the methylation level of a given gene promoter should be correlated with the level of its gene expression? Then we can say that methylation regulates its expression. In...
01 January 1970 6,015 3 View
I would like to study epigenetic regulations in the reproductive system of cows. I have already done the initial research and would like to continue it in a larger grant project. However, I met...
01 January 1970 3,898 0 View
Dear scientists. Have any of you published the results of changes in gene expression obtained by next-generation sequencing? The analysis was carried out in endothelial cells of cows with some...
01 January 1970 6,231 2 View
