Does any one have any protocol or assays by which I could study signalling profile of tor gene during various stress conditions in yeast?
Hello. I want to crystallize kinase domain of a protein in active form. The active form of the domain has specifically phosphorylated tyrosine residues. However, I am not sure if I can do so in a...
01 February 2020 7,569 5 View
I have purified a protein, however I am getting double band on SDS-PAGE. Even after FPLC the double band doesn't resolve and become more prominent. I have checked through Mass spec that both the...
06 July 2019 6,753 10 View
Is their any online bioinformatic tool available through which I can understand or know the interacting partners of a kinase domain of the protein I am studying? I wish to study interactome of...
06 July 2019 6,134 6 View
I am purifying a protein expressed in pLysS cells. The construct is in pET28 vector and having His tag with TEV cleavage site . Now when I ran the SDS-PAGE gels after TEV cleavage and...
03 April 2019 7,604 16 View
In my construct I have two His tags, one at N-term and another at C-term. There is a TEV site between the N-term His tag and my gene but no cleavage site for the C-term. So, do I need to add...
03 April 2019 2,343 8 View
Is it that vectors with T7 promoter do not grow/transform well in rosetta or codon cells?
02 March 2019 1,036 3 View
I am not getting colonies when I had transformed my construct in rosetta 2 cells whereas transformation worked in Bl21-pLyse cells. I had used the same heat shock protocol for both. The one change...
01 February 2019 2,244 11 View
Can any one provide me appropriate protocol for preparing crude mouse tissue lysate for western blotting. The protein of interest resides both in cytoplasm and nucleus so which lysis buffer would...
01 February 2017 5,391 6 View
I want to study genetic relatedness among fresh water fishes, so which molecular marker would be best for this study?
07 August 2016 6,257 2 View
I want to test expression of one of the protein in clinical samples of oral cancer. So can I use cheek cells as control or I need to derive some other tissue from mouth of normal individual?
06 July 2016 1,360 4 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 all! My thesis groupmates and I are working on purifying an E. coli bacteriophage. The phage was handed down to us by the previous thesis group, but they only managed to purify it to about...
06 August 2024 5,801 4 View
I have prepared a liquid broth containing Yeast (S. Cerevisiae) that i need to add from it to fermentation media of lignocellulosic hydrolysate. I need to know based on what parameters do we add...
06 August 2024 662 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 tried several times to isolate lymphocytes from mouse spleen, but all attempts have been unsuccessful. I tried most available protocols. I used different dissociation media (HBSS with Ca...
04 August 2024 9,913 7 View
What is the role of oxidative stress in toxin-induced carcinogenesis? I am currently researching the impact of environmental toxins on children's health and would greatly appreciate insights from...
02 August 2024 3,174 1 View
I have been attempting to extract DNA from Bacterial, Fungal and Yeast banked samples (>1e7 cells) using Prepman Ultra reagent and I seem to be struggling to obtain a sequence. Although the...
01 August 2024 2,079 0 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