T7 promoter and Rosetta?

More Shubhashish Chakraborty's questions See All

Why is there cell debris are coming after 2nd or 3rd passage of the cells???

Our lab is facing a difficult problem for almost a year. After thawing the cells are okay, but after 2nd or 3rd passage there are debris present between the cells. We use mycoplasma removed cells...

17 January 2021 5,629 6 View

Can you help me for generating chicken swarm optimization matlab code?

Multi-objective optimization through chicken swarm optimization technique. Both maximization and minimization problem.

31 December 2020 2,299 4 View

Any good methods for double-stained markers quantifications from tissue sample using computer programs?

How can we best quantify double-stained markers when comparing diseased and control? For example, if we want to stain one marker of smooth muscle cells and compare the expression in diseased and...

21 December 2020 7,389 1 View

Membrane protein Expression Purification: Broad Asymmetric peak on size exclusion ?

Hello, I am expressing an integral membrane protein (MW 51kDa) in Hi5 insect cells. I initially purified the protein by Nickel-His, concentrated it and subjected it to Size Exclusion...

10 December 2020 2,781 5 View

How should I store the milk sample in RNAlater?

I want to extract RNA from human breast milk. How should I store the milk sample in RNAlater? And after RNA extraction how can I store the RNA?

01 December 2020 7,183 4 View

Can U suggest standard protocols of quantifying total phenolics & total flavonoids using gallic or tannic acid for the former and quercetin for later?

The problem is that everytime getting the flavonoids content much higher than phenolics which may be, but in rare occasion. In phenolics, I am adding100 ul sample with 900 ul dH2O with...

29 September 2020 4,993 6 View

What is the ideal Elution buffer composition for a His tag protein purification to prevent protein aggregation after freeze thaw ?

Hi All, Recently, I purified 8 proteins and stored at -20 with the elution buffer containing 20 mM Tris, 250 mM NaCl and 200 mM Imidazole. I autoclaved the buffer, so I do not know whether the...

25 September 2020 9,814 6 View

What is the use of having multi-wavelength absorption measurements in Aethalometer?

Aethalometers are used to estimate the light-absorbing aerosols or Black Carbon (BC) or Soot particles. There are widely used aethalometers which are measuring light absorption in multiple...

06 September 2020 1,050 3 View

Has anyone done magnetic target localization using Genetic Algorithm?

I am trying to model magnetic target localization in MATLAB using those algorithms. So essentially what I want to output is the position vector of the target (x, y, z) and the magnetic moment...

28 August 2020 9,811 2 View

Can an isolated protein domain give a single band in SDS-PAGE equivalent to its dimeric mol. wt. ?

The protein I'm working with forms a homodimer and is a 2-domain protein. I have cloned and purified one of its domains. However, when I run the domain on SDS-PAGE, I get a single band equivalent...

19 August 2020 7,893 14 View

Can linear DNA be used as template for site-directed mutagenesis?

Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...

03 March 2021 401 4 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 View

Runs of nucleotides in Sanger sequencing?

I performed site directed mutagenesis, transformation, and then I sent out plasmids for Sanger sequencing and found out that there is extension of DNA just before the stop codon. I am not sure...

27 February 2021 547 3 View

I want to performe MALDI-TOF MS for bacterial identification, I would appreciate it if anyone can tell me the matrix solution preparation protocol ?

based on what I've studied on multiple papers, for peptide and protein identification Alpha-cyano-4-hydroxycinnamic acid (CHCA) is an appropriate matrix to use, I also know that the CHCA powder...

26 February 2021 6,799 3 View

Difficulty in cloning of 1kb gene into vector?

I am facing difficulties in cloning a 1kb gene into a vector (pJIT163). I have my gene on interest (GOI) in pUC57 and want to clone in pJIT163 using SalI and BamHI restriction sites. I am getting...

25 February 2021 3,221 7 View

Universal bacterial primers targeting a mRNA?

Hi, I'm looking for universal primers targeting a bacterial mRNA to quatify bacterial load by qPCR in a complexe mixture (eukaryote and prokaryote RNA). Ribosomal RNA being depleted, I absolutly...

22 February 2021 9,969 3 View

Covid 19 Pandemic ' impact on digital transformation?

Covid 19 Pandemic and digital transformation

22 February 2021 3,351 5 View

Does anyone have any ideas why drug-resistant malaria is increasing in Southeast Asia even when those drugs are no longer used?

When dealing with bacterial toxins, there are three varieties of microbes that develop in your petri dish: toxic, resistant, and susceptible. Initially, there are only toxic and vulnerable...

21 February 2021 6,084 5 View

I can detect my protein but the encoding DNA sequence has stop codons within. Should I ignore the stop codons?

I constructed a synthetic DNA library (scFv, VH-VL orientation) with a 3' reporter and 8x histag. I cloned and expressed this gene following which I performed an ELISA. The ELISA results suggested...

19 February 2021 7,006 5 View