How to avoid protein precipitaion ?

I am working on a thermophilic protein (pI: 7.72) purified using Ni-NTA chromatography (Final buffer: 25mM Pipes (6.6), 300mM NaCl, 1mM PMSF, 10% glycerol, 200mM imidazole). But the protein...

09 October 2016 8,241 24 View

Does cloning vectors (pET series specially) confers toxicity to the gene/protein of interest?

I am working on a thermophilic protein which is not known to be toxic. It was earlier cloned in pET11a and cell death was observed after 2-3 hrs of IPTG (0.1mM) induction. I then cloned the gene...

06 July 2015 751 4 View

Why do I detect two proteins after purification?

While eluting two of my proteins with 250mM of Imidazole, I am getting two different bands (Size of the proteins are mentioned in the Image attached). The genes are cloned in pET22b without Signal...

06 July 2015 1,710 8 View

Is it recommended to incubate Thermophilic genes at 42 degree celsius after IPTG induction?

11 December 2014 3,428 5 View

Why are my non-specific bands coming even after PCR clean up?

I am trying to clone a T.thermophilus gene. I have used 3%/5% DMSO during PCR amplification (using phusion enzyme). Amplification is coming but after PCR purification, and I am also getting...

05 June 2014 2,160 21 View

Why is the ligation of a gene with different vectors not coming?

I have tried cloning of my gene of interest in pET22b and pET28a, for different ratio, but the ligation didn't work (though colonies are coming in some cases). I am pretty much sure that my PCR...

04 May 2014 6,595 14 View

Do you have any suggestions on how to avoid primer dimers after PCR clean up?

When I am doing PCR clean up after PCR amplification (using phusion + DMSO) of a thermophilus gene, and along with the amplified gene, the primer dimer is also appearing. Do you have any...

03 April 2014 1,399 18 View

What is the denaturation temperature used for plasmid vectors (circularised) in PCR? Is it different from 95 degrees C?

I am amplifying a gene from pET11a vector. Is the temperature for denaturation (95 C for chromosomal DNA generally) different for amplifying the gene from plasmid DNA?

02 March 2014 6,215 5 View

Is there any protein expression host designed for thermophilic bacteria? Can it be expressed in E.coli hosts namely Rosetta, C43 DE3 or not?

I am trying to purify a protein from a thermophilic bacteria (gene cloned in DH5 alpha) but I am not getting expression of my protein after IPTG induction. I have tried with different expression...

02 March 2014 2,976 11 View

Is alkaline picrate method considered a less sensitive method for creatinine estimation?

What are the other substances which can be used that give a positive result with alkaline picrate?

10 November 2013 8,819 3 View

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02 March 2021 3,060 3 View

What do you know about biotechnology art?

What do you know about biotechnology art?

02 March 2021 1,653 3 View

Which is the best place in the world for the plant biotechnology research?

I am searching for a good place for the Post Doc,

02 March 2021 4,053 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 View

How was your experience with Biomedical and Biotechnology Journal?

How was your publication experience with BBRJ in terms of peer review and reviewer comments. Did you find it fair and transparent ?

23 February 2021 3,810 6 View

What are the best cations for preparing competent cells, Ca2+ or Mg2+?

I have been reading various articles and most state that treatment with CaCl2 produces higher efficiency of transformation than MgCl2. Can anyone explain why Ca2+ is better than Mg2+ at...

16 February 2021 8,519 5 View

Kinetics of Enzyme production in Recombinant E Coli?

Hello, I am trying to find kinetic information (such as rate of enzyme production etc.) on the expression of taq polymerase in E-Coli after inserting a recombinant plasmid. Any help would be...

12 February 2021 1,386 3 View

Are the topics below viable research in plant biotechnology?

1. Chemically induced mutagenesis in seed and qPCR detection and amplification of desired trait. 2. Site-directed mutagenesis in qualitative traits of a multiline seed.

04 February 2021 9,069 3 View

Is receiving Invitation to submit a article from the journal is big deal ???

I have recently received a invitation from the journal "Current Pharmaceutical Bio-technology" to submit a research article. As i didn't received any invitation before. so, is it normal to to...

04 February 2021 4,596 23 View

Alternatives to .45 um filter to isolate MS2 from lysed E. coli?

Hello, I am looking for alternatives or cheaper materials to isolate out MS2 from E. coli following lysis via chloroform. Some literature I have reviewed uses a .45 um nanopore filter following...

02 February 2021 807 3 View