16 Questions 24 Answers 0 Followers
Questions related from Stefan Paulusch
hello, i am planning to transfect mouse primary hepatocytes with miRNA in order to isolate microsomes to study the activity of my enzyme of interest. Therefore, I wondered if the primary...
08 August 2019 2,113 2 View
Hello, how can I calculate the actual amount (in mg) of beta-glucuronidase from supplied white powder lyophilized from 0.1 M sucrose plus 1 mg sucrose per mg protein ? On the delivered vial it is...
10 October 2018 4,809 0 View
Hello RG community, My task is to measure the expression of several microRNAs from total RNA isolated from human body sera. I have a number of TIPS patient (suffering from liver cirrhosis) and...
08 August 2018 5,185 1 View
Hi there, I received extracted whole RNA from TIPS patient serum and negative healthy controls. All samples contained spiked-in SV40 miRNA already as a reference gene. I performed miRNA reverse...
04 April 2018 4,006 5 View
Hello RG community, I want to determine the effect of miRNAs on the 3´UTR of retinoid X receptor alpha mRNA and therefore want to analyze its mRNA level in HepG2 cells. I want to go and check for...
03 March 2018 1,101 5 View
Hi guys, I have a question about glucuronide CDCA-24G, maybe someone has already worked with it. I want to perform an experiment in hepatocytes which I have extracted from mouse livers. I do not...
02 February 2018 1,892 2 View
Hi all, I tried to insert a restriction site in a 6.5 kb plasmid having only six endogenous restriction sites in the MCS. I already tried several primer setups but all failed. What could I...
11 November 2017 3,539 10 View
Hi guys, a situation has just come up. I want to integrate a restriction site into a Plasmid (~6.5 kb) by PCR mutagenesis. After PCR (25 cycles), I observed the DNA band on the gel, however very...
09 September 2017 8,214 4 View
Hi all, I would like to use pMIR-REPORT for cloning of the 3´UTR of interest and study the interactions of exogenous miRNAs but this plasmid is only available in a kit from Thermo which is quite...
08 August 2017 2,005 4 View
Hi all, does someone have experience with studying the interaction of microRNA (miRNA) and its target sequence cloned into a Luciferase Reporter vector? Actually using pGl3-Basic vector from...
08 August 2017 3,577 5 View
Hello all, I wondered whether the equation is right that we use in the lab for HepG2 cell counting with the Neubauer counting chamber. We generelly use a sample volume between 40-50 µL of cells...
09 September 2016 8,961 2 View
I got my HEPG2 cells passaged to a new cell culture flask and have mistakenly used 10 X PBS for washing instead of 1 X PBS. Although washing took relatively short time how bad might be the...
06 June 2016 5,661 3 View
Hello dear community, I am about to start Investigating transcription Regulation with miRNA and I am particularly interested in the question how miRNA come to upregulate gene Expression. Does...
06 June 2016 7,114 6 View
A colleague of mine has some diffuculty with his RT-PCR experiment. He started analyzing the single failure-prone steps telling that he has restreaked the strain of interest due to some growth...
10 October 2015 3,651 8 View
Hello everyone, since my supervisor left already I am desperately looking for the restriction map of plasmid pMHH1. I want to start curing my E. coli of its plasmid by using suicide vector pMHH1....
07 July 2015 5,366 7 View
Since freeware like PlasMapper annotate genomes up to 20 kbp I could not find a reliable tool for a graphical analysis of my 80 kB plasmid. I performed RAST annotation already which was quite...
07 July 2015 8,698 4 View
