15 Questions 18 Answers 0 Followers
Questions related from Stefan Paulusch
Hello RG community, I wonderer whether exogenous molecules (e.g. reporter plasmids, miRNAs or other artificial molecules) are transferred between cultured human cells during their doubling ? So,...
22 February 2021 7,027 3 View
Hi all, I just listened to a discussion about protein domains and a potential substrate binding. A question was given and I was thinking how to tackle the problem. Let's say: a functional protein...
19 June 2020 1,425 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...
31 August 2018 9,522 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...
09 April 2018 5,627 2 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...
28 February 2018 5,195 2 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...
28 September 2017 2,737 3 View
Hi RG community, we started cultivation of Hek293 cells in DMEM GlutaMAX from Gibco plus 10 % FCS and Pen Strep. The cells grow pretty slowly presumably due to the medium composition. We´re...
06 July 2017 2,436 8 View
Hi, I´m working on site-directed mutagenesis trying to mutate several short stretches of ~6-7 nt within a 5 kbp plasmid. I designed mutagenesis primers. In 2 PCR reactions the 2 DNA fragments are...
02 May 2017 2,654 3 View
Hi, two days ago I prepared competent E. coli JM109 by a protocol using 0.1 M CaCl2 solution. I grew the culture to an OD600 of ~ 0.5, spun them down and resuspended in chilled 0.1 M CaCl2. After...
28 April 2017 8,857 4 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...
29 June 2016 3,627 4 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...
21 June 2016 9,558 3 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...
26 October 2015 3,181 7 View
Unfortunately, there is no centrifuge in our lab spinning at 20,000 x g so I wondered whether I can just use our centrifuge at Maximum rcf ( 5000 x g) and spin the sample by doubling of the time...
14 July 2015 8,511 1 View
Hi all, For my plasmid isolation the protocol says to use 2 M acetic acid but our store has only got 100 % pure glacial acetic acid. So how can I make a 2 M acetic acid? Happy for any help!
02 July 2015 3,731 12 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....
01 July 2015 1,608 7 View
