Bharat Bhusan Patnaik

10 Questions 44 Answers 0 Followers

Questions related from Bharat Bhusan Patnaik

What are standard PCR conditions for amplifying a 3 kbp DNA fragment?

I have been trying to amplify two genes (basically the ORF region) that have sizes in the range of ~3000 bp for subsequent cloning and sequencing. I am initially using pfu polymerase for...

01 January 2014 4,467 13 View

Why is there a difference in the genomic length assessed by electrophoresis and calculated by sequencing?

We have worked out the whole genome sequence of a virus by the traditional sequencing methods (shotgun library). The size of the circular-form of the genome after sequencing is 108kb, whereas...

10 October 2013 3,578 2 View

What is the reason for getting a poor yield of dsRNA for gene silencing experiments after in vitro transcription process?

My work relates towards knockdown of an important gene in the Toll signaling pathway and study the effects on AMP production. I started off with the gene contained in plasmid and got a good...

04 April 2013 9,857 11 View

Can we patent genes (full-length cDNA) deciphered by a novel strategy?

I have deciphered the full-length cDNA of a plant-specific gene by a novel strategy, not reported before. The full-length cDNA is also novel as it hasn't been reported before in literature. I...

07 July 2012 8,778 11 View

Cloned insert (TA Cloning vector) in reverse orientation

I have been conducting TA cloning of my gene of interest for its expression but mostly I get sequencing results of the clone inserted in reverse orientation rather than the correct orientation....

06 June 2012 9,871 6 View

Double digestion of TOPO pCR 2.1 TA Cloning vector with the gene of interest not giving insert band of expected size

My gene of interest (Partial N-terminal fragment) have been cloned in TOPO TA Cloning vector-Invitrogen. I want to express the gene in the expression pET vector and produce the recombinant...

05 May 2012 6,934 12 View

Novel sequence submission in public databases

I have sequenced (currently partial nucleotide sequence) and would like to submit the same to the gene and protein databases. I have already posted the N-terminal protein sequence to Uniprot and...

04 April 2012 5,313 8 View

Detection of protein complexing in Native-PAGE of purified bioactive protein.

I have purified a novel bioactive protein and have characterized the same using SDS-PAGE, intact mass determination by MALDI-TOF, Peptide mass fingerprinting and Edman sequencing to be of 21.5 kDa...

03 March 2012 8,108 7 View

I have purified my bioactive proteins by RP-HPLC (linear gradient flow of acetonitrile with water) with two peak fractions at 280nm absorbance. I have...

HPLC purified peak fractions lyophilized and run on 12% SDS-PAGE- No band observed with coommassie staining. Please suggest.

02 February 2012 3,487 6 View

How to study the activity of a purified protein/peptide against viruses?

Precipitation test in microwell plates

12 December 2011 2,782 7 View