People I need some help here.
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
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.
09 October 2014 6,179 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
My name is Apurva Saoji. I am a Ph.D scholar in Computer engineering in India. I am looking for international expert in reviewing my PhD thesis, "Competitive Optimization Techniques to Minimize...
07 August 2024 4,600 2 View
The political participation of the Kirat Rai community in Nepal has evolved significantly over time: Historical Context Historically, the Kirat Rai people, like many indigenous groups in Nepal,...
05 August 2024 5,950 1 View
I aim to be as skeptical as possible regarding whether a pair of orthologous genes results in the same phenotype in their different but related bacterial organisms under similar environmental...
05 August 2024 6,787 4 View
I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....
05 August 2024 2,570 3 View
I have an RNA-seq data that I have analysed using Limma-voom and have extracted the gene IDs, log2FC and the p-values. At p value < 0.05, I have over 10,000 DEGs, however, when I run the GO...
31 July 2024 225 2 View
I am currently working on a project involving liposomes and need to determine the maximum volume of siRNA that can be added to a 2.5 mL liposome solution with a total lipid concentration of 10...
30 July 2024 6,420 1 View
I cannot confirm cell surface protein expression by flow cytometry even after transfection and antibiotic selection of cells. Does it take long for proteins to express on the cell surface? the...
28 July 2024 3,178 3 View
I created two potential gene expression cassettes (constitutive and inducible) for expression of a mutant PETase gene on PeptiCloud using the version tree feature, which allows users to create...
28 July 2024 7,559 1 View
How much volume of siRNA can I add to 2.5 mL of liposomes with a total lipid concentration of 10 mg/mL?
27 July 2024 6,920 1 View
Gene sequencing related trouble shooting
25 July 2024 4,149 2 View
