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

More Suraj Kumar Mandal's questions See All

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 8,338 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 803 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 1,772 8 View

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

In some papers people have recommended to use incubation at 42 degree celsius for short time for protein expression after IPTG induction? If YES, What will be its effect on protein's...

11 December 2014 3,484 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 2,234 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 6,663 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 1,451 18 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 3,023 11 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 6,313 5 View

Which part of the alsA gene in E.coli is bound to the transmembrane domain?

alsA gene has two sub-units and some part of one sub-unit is bound to the TMD (alsC).

10 November 2013 9,698 0 View

What does negative readings in spectrophotometer denote. Was testing TSS content of bacterial cultures?

I was performing estimation of Total Soluble Sugar content in bacterial cultures by Anthrone method. Out of 9 samples, only one sample showed positive reading while others had a negative reading...

22 July 2024 9,576 4 View

My article: The Extended Pedersen hypothesis can you correct the the name of first Author?

In My article: The Extended Pedersen hypothesis (published, February 1988 Clinical Physiology and Biochemistry 6(2):68-73,) there is mistake of the name of first Author: it is not Catherine...

11 July 2024 1,801 2 View

Is it possible to generate a (semi-)permanent "transcription bubble" within a replicable plasmid?

I was wondering if it is possible to form a permanent open "ssDNA bubble" similar to a transcription bubble (>13 nucleotides) within E. coli. These criteria are important: 1. Open ssDNA...

07 July 2024 8,275 5 View

How to dilute essential oil from plants to assess the biological activity?

I have extracted essential oil from a medicinal plant and want to make several concentrations for some biological activities (Antidiabetic assay). How can i prepare the several concentrations to...

04 July 2024 8,100 4 View

Nutrition map: What does it mean for you?

Unluckily, just similar to an expensive car, your brain can be damaged if you ingest anything other than premium fuel.

23 June 2024 2,287 7 View

Have pathomorphological studies of the lungs been performed in patients with desminopathy?

It is known that patients with desminopathy often die from pneumonia. Have pathomorphological studies of the lungs been performed in patients with desminopathy?

15 June 2024 1,355 7 View

Luciferase assay lysis?

Hi! I'm doing a renilla luciferase assay with coelenterazine and HEK293T cells transfected with AP-1 renilla luciferase. I'm wondering if the cells need to be lysed prior to adding coelenterazine...

13 June 2024 9,936 5 View

How to prepare 0.28 U/ml working solution of Acetylcholinesterase inhibition assay for anti-Alzheimer activity?

I have purchased Acetylcholiesterase from Electrophus electricus (electric eel), C3389-2KU, following details are written on the enzyme vial. Type-VI S lyophilized powder 200-1000 Units/mg...

31 May 2024 6,444 1 View

What is the difference between absorption and adsorption?

30 May 2024 4,447 27 View

What effect may biochar types produced from grape stems, leaves, onion leaves and stems, asparragus and cantaloupe stems/leaves have on a harvest?

I'm interested in knowing the differences in the effects caused by four different types of biochar: cantaloupe leaves/stems, onion leaves/stems, grape peels/stems/leaves and asparragus...

28 May 2024 4,907 0 View