02 February 2013 0 9K Report

blaLEN and bla SHV are a class of beta lactamases that has resulted in evolution of drug resistance in Klebsiella world wide.

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Extra DNA band in DpnI based point mutation?

There's a clone in pBAD18cm vector (6800bp) which was subjected to point mutation through PCR-based technique. After PCR and DpnI digestion an extra band of approx 1500bp was observed in cloned...

01 February 2016 4,561 4 View

What are the best free softwares to study and analyse enzyme kinetics data?

I am doing enzyme kinetics studies. I need to derive certain parameters for a representable data. The present MS excel sheet fails to provide enzyme kinetics parameters such as Km, Kcat, Vmax...

10 November 2015 7,277 7 View

What is the best solvent for carbapenem antiobiotics? Water or DMSO?

A very basic query regarding the solubility, stability and storage of carbapenems antibiotics such as Meropenem trihydrate or Imipenem monohydrate. I have known to dissolve them in DMSO in higher...

09 October 2015 5,403 8 View

Are there any ghost bands in my SDS-PAGE gel or contaminating proteins?

Here I have attached SDS-PAGE gel picture of Ni-NTA affinity chromatography elution profile of a protein at varying concentration of imidazole in tris buffer(pH 8). I have done washing in 10, 25...

07 August 2015 1,667 8 View

How do I get higher molecular weight band after PCR clean up followed by gel extraction of double digested inserts?

It's confusing to find extra bands of higher molecular weight in my insert sample which was PCR purified by gel extraction method. The insert bands were very clean in 0.8% agarose gel. It was...

05 June 2015 4,240 13 View

Is it possible to get a protein of interest at higher molecular weight than expected in SDS-PAGE?

I am checking in-vivo expression of a protein(full length) in pBAD18cm vector using BL21 expression host. I am getting an extra band of protein, 10kDa higher than expected in SDS-PAGE as compared...

04 May 2015 6,479 10 View

What are the good servers for detecting signal peptides apart from SignalP 4.1 and Signal blast?

What are the good servers for detecting signal peptides apart from SignalP 4.1 and Signal blast? Sometimes these servers fail to detect the presence of signal peptide in a given protein sequence....

04 May 2015 8,435 2 View

Does NDM-1 have signal sequence?

I tried to check the presence of signal sequence of full length construct (amino acid) of NDM-1 using SignalP 4.1 server but could not find it. Can any body suggest anything in this matter? Thanks

04 May 2015 8,397 0 View

Why there are two bands in SDS-PAGE of a purified protein instead of one?

I have cloned a beta-lactamase gene in BL21 E.coli strain using pET28a vector for expression and in vitro studies. After Ni-NTA column purification of this His Tagged protein, I am getting two...

31 December 2014 2,330 16 View

What should be the ideal dialysis buffer composition for a His-tagged purified protein to remove imidazole salts?

My His-tagged purified protein contains 10mM trisCl, 300mM NaCl and 200mM imidazole at which the protein was eluted at pH 8.0. What should be ideal dialysis buffer composition to remove...

31 December 2014 5,867 10 View

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