16 Questions 25 Answers 0 Followers
Questions related from Shanice Chiang
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.
10 October 2014 6,145 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...
10 October 2014 3,654 3 View
People I need some help here.
10 October 2014 8,802 0 View
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...
10 October 2014 8,647 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.
10 October 2014 9,421 4 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?
09 September 2014 5,403 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...
09 September 2014 5,525 4 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...
08 August 2014 3,758 13 View
what is the most effective method here
08 August 2014 8,383 2 View
Dear all, I tried to extract RNA from different rice plant tissues and then convert it to cDNA. I then amplified the cDNA with different set of primers. Only one of them showed the expected size....
07 July 2014 3,043 11 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...
07 July 2014 1,807 4 View
Is there any other ways of testing the RNA integrity without using bioanalyzer or denaturing gel? Has anyone tried bleach gel before? I tried it but it doesn't work well. Wondering if anyone is...
07 July 2014 6,839 5 View
I have cloned a rice gene into the pET28a His tag vector. I have confirmed recombinance by sequencing and it is in frame. I run the SDS PAGE along with my empty pET. I am attaching my gel...
07 July 2014 6,372 12 View
Hi all, I have just cloned my gene into pET and would like to check if my gene is in frame. Can I just translate my sequence and then do a protein blast is that a proper way of doing it?
07 July 2014 5,564 4 View
Dear All, I need some help. I have been trying to clone my gene into pET28+ for months but the results is always disappointing. My brief protocols are as per follow: 1. PCR on my gene of...
07 July 2014 2,107 18 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)...
07 July 2014 1,541 6 View
