I am not sure if my induction have worked. Is it possible to do a RNA miniprep. IF RNA successful extracted- means induction has worked.
Hi people, I am having a lot of difficulties in my protein expression. I am wondering if my induction has failed to work? Too bad, i dont have any positive control. So i am using the empty...
09 October 2014 8,665 9 View
I am going to do induction in Pichia pastoris. But my lab does not have yeast nitrogen base. Just wondering if I can substitute with some other things.
09 October 2014 9,470 4 View
People I need some help here.
09 October 2014 8,822 0 View
Do you know of any website which would predict RNAse E cutting site? or how to predict it? We are having problems expressing a foreign mRNA in E.coli and one possibility was that the mRNA was...
09 October 2014 3,671 3 View
I need lactose for my autoinduction media for E.coli. Just wondering if I could use the D-lactose monohydrate because I can't find any alpha-lactose monohydrate in my lab. Are they the same?
08 September 2014 5,424 8 View
I have cloned lipase gene into pET28+ and express it in Rosetta(DE3)pLyss. I have tried to induce with IPTG but I can't spot any difference between the clone and the empty pET. I varied the...
08 September 2014 5,549 4 View
what is the most effective method here
07 August 2014 8,404 2 View
Dramatic induction was observed, but no desired protein band was observed. I run an empty pET as a control. There doesnt seem to be difference with the empty pET and pET with insert. I have no...
07 August 2014 3,782 13 View
I have cloned my lipase gene from rice in pET-28(+) His-tag vector. Was wondering how do I choose/design the primary and secondary antibodies for western blot. I am totally new to this. Was hoping...
06 July 2014 1,835 4 View
Protein Expression in Prokayotic (Rosetta (DE3) E. coli) (1) Transformant Rosetta (DE3) cells with recombinant pET28(+) were grown on LB/Kanamycin/Chloramphenicol agar plates overnight. (2)...
06 July 2014 1,558 6 View
Hello, I am currently having problems with RNA extraction. I am using mouse liver (C57BL6J), and I have extracted RNA from mouse liver before. Before this experiment, my final RNA pellets were...
11 August 2024 7,082 3 View
I have been doing the m6A dot blot for a while with no improvement, I am extracting the RNA, and I can see the dots although the three biological replicas give a different reading on the memberan...
10 August 2024 8,539 5 View
I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...
08 August 2024 4,645 2 View
I've been performing RNA extraction on cotton petiole tissue for a few months now using the method described in the following paper, a derivative of the typical hot borate method...
08 August 2024 9,882 2 View
After performing symmetric PCR, PCR purification was performed. Afterwards, asymmetric PCR was performed using the PCR purification product as a template, but no ssDNA band was confirmed in the...
08 August 2024 1,668 3 View
I am currently working on LncRNA; to know the lncRNA-protein interactions I want to do RNA pull down assay, so I need to design primers with T7 promoter. I need assistance in this regard.
07 August 2024 6,622 1 View
Hi! So i attempted to understand a novel protein behavior towards heat application by analyzing its secondary structure change. I subjected the protein to a thermal denaturation analysis using...
06 August 2024 1,989 3 View
Recently, we observed that 99% of the sequences in our RNA-seq data corresponded to the E. coli genome. Despite multiple DNAse treatments after RNA extraction and ribosomal depletion, we were...
06 August 2024 807 3 View
I am planning to collect human fecal samples for metatranscriptomic analysis using MGI. These samples are from indigenous people living in a region with high temperatures. I will have access to a...
06 August 2024 1,367 3 View
i have sorted anti-NP specific plasma cells from bone marrow of C57BL/6 mice at certain times after immunization with variable counts and isolated total RNA using TRIZOL method for RT-PCR using...
05 August 2024 8,835 1 View