T7 promoter and Rosetta?

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Phosphorylation of kinase domain in bacterial expression system?

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

Double band after protein purification?

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

Tool to understand the interactome of a kinase domain of a protein?

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

Double bands in SDS-PAGE gel?

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

No cleavage site in C terminal having His tag?

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

Transformation in Rosetta2?

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

Protocol for lysate preparation from mouse tissues for western blotting ?

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

Marker for determining genetic relatedness and distance?

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

Control for oral cancer?

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

Assays or protocol for TORC signalling?

Does any one have any protocol or assays by which I could study signalling profile of tor gene during various stress conditions in yeast?

06 July 2016 9,976 0 View

How to confirm the site-directed mutagenesis result without performing NGS?

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

Is there any way to quantify bacterial and fungal cells in their mixed culture?

I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...

07 August 2024 7,535 4 View

How can we now if the promotor in the vector is well?

Hi, everyone. I want to transfer two genes in to the pseudomonas bacteria that isolated from the soil. The bacteria that I want to use for cloning haven't been identified and not be sequenced,...

02 August 2024 3,987 1 View

How is the bacterial genome's high protein count verified as genuine despite 800+ contigs and good metrics (98.55%completeness, 0.68% contamination)?

Given that the bacterial genome has over 800 contigs, but its quality metrics are good, with a completeness of 98.55% and a contamination of 0.68% as assessed by CheckM, what specific validation...

01 August 2024 1,514 1 View

How to back transform the results generated from analyses using log transformed with In(X+1) data?

I am conducting my analysis using SPSS. I log transformed my data using In(X+1) as my data contain zero values. However, when I want to back transform the regression coefficients generated from my...

31 July 2024 7,860 3 View

Is it possible to use a safer acid instead of monochloroacetic acid to convert bacterial cellulose to carboxymethyl cellulose?

Hi, researchers, Is it possible to use a safer acid instead of monochloroacetic acid to convert bacterial cellulose to carboxymethyl cellulose?

29 July 2024 4,570 0 View

Recovery Viurses from bacteria genome?

Hello everyone, I am currently looking for tools to recovery viral genomes from bacterial genomes, not metagenomes. However, I have only found tools that are designed for retrieving and studying...

28 July 2024 8,953 1 View

Question about MHB (Mycorrhiza Helper Bacteria) isolation technique??

What are the most effective isolation techniques for obtaining MHB?? Is there are best approach techniques? I keep struggling in obtaining it.

26 July 2024 7,387 0 View

Illustra™ MicroSpin™ G-25 columns what it is used for?

Hello, We found three packages of Illustra™ MicroSpin™ G-25 columns in the cabinet of an unused lab. They are very old but have never been opened. I have never used this kit before, and I couldn't...

25 July 2024 4,927 3 View

Swerling Characteristic functions?

Hello!!! I want to implement the Swerling characteristics functions (CF) directly in MATLAB without using its Fourier integral pairs...the Swerling CFs are actually Laplace Transform of the signal...

23 July 2024 4,925 1 View