Atul Sathe

9 Questions 10 Answers 0 Followers

Questions related from Atul Sathe

Will CRISPR-Cas9 knockout of a gene lead to reduction in the level of it's expression?

Generally, CRISPR-Cas9 editing leads to the formation of indels at the target site in the coding sequence of a gene. This often results in an altered reading frame (frameshift mutation) which will...

09 January 2025 5,051 2 View

How to get the correct transcript out of alternatively spliced transcripts?

I'm trying to clone the CDS of a gene. This gene harbors 12 Exons and 11 Introns. Size of the CDS is about 4 kb. After amplifying the CDS, I have cloned it into a cloning vector. Sequencing result...

07 August 2017 3,619 10 View

How far the promoter is located from the start codon (ATG) in Eukaryote genome?

I want to amplify the promoter region for one cloned gene in rice. For this I'm going to amplify 2 kb region (from genomic DNA) before the start codon (ATG). I have a concern regarding the reverse...

10 April 2017 1,216 6 View

DNA sequencing result shows presence of deletion, but at cDNA level it shows there is no mutation, why this so?

Im working on rice mutant. After mapping and sequencing the gene, I found that there is a 6 base deletion occurred in the gene. But when I tried to PCR amplify using cDNA, the amplicon size is...

20 October 2016 4,357 3 View

How can we overcome seed borne bacterial contamination in cotton?

We are working on plant tissue culture in cotton. The present bacterial contamination is seed borne (Gram -ve, cocci, in chains). We have tried to overcome it with 0.1% HgCl2, 0.1% SDS as well as...

03 March 2015 4,884 8 View

How to reduce or eliminate the non-specific amplification in PCR?

I'm using the gene specific primer for screening the plant DNA samples. At the specific annealing temp (55) a 500 bp amplicon is getting amplified but there are significant non-specific bands too....

03 June 2014 6,439 12 View

What is meant by Chimera/Chimeric transformation?

In case of genetic transformation (either Agrobacterium mediated or by particle bombardment), we will get transformed plant cells which may be either stably transformed or chimeric transformed...

11 February 2014 6,529 8 View

How to control Agrobacterium outgrowth in case of cultured cotton explants?

Usually we do co-cultivation for 2 days followed by transfer to plain media where Agrobacterium outgrowth starts to appear and goes on increasing. Even if we have treated contaminated (with...

26 November 2013 1,145 4 View

What is the appropriate or most widely used concentration of Acetosyringone for Agrobacterium mediated plant transformation?

Is there any alternative to ACS?

15 August 2013 6,739 1 View