Anil Kumar

5 Questions 12 Answers 0 Followers

Questions related from Anil Kumar

Kindly suggest me the ways to improve the transfer efficiency of DNA on to positively charged nylon membrane for southern hybridization?

I am working on identification of copy number in a recombinant Chinese hamster cell line by southern hybridization. I am having issues with the transfer of DNA onto positively charged nylon...

09 February 2024 2,696 8 View

What is the ideal restriction digestion reaction volume for southern blotting? Can we scale up the volume to 200 ul if gDNA concentration is low?

while working with low concentrated mammalian genomic DNA, can we increase the restriction digestion reaction volume to 200 ul to accommodate the required amount of mammalian genomic DNA for...

23 January 2024 7,002 12 View

Does width of the slot in the gel have any effect on the transfer efficiency and signal detection sensitivity in southern blotting?

While preparing the gel for resolving the restriction digested fragments of mammalian genomic DNA and subsequent transfer on to the nylon membrane, is it very critical to consider the width of...

23 January 2024 4,372 0 View

Can anybody suggest me a protocol for RNA extraction from the least possible amount of sample (approx. 30- 50 mg) of developing Arabidopsis siliques?

I am working on a time course expression analysis experiment with developing Arabidopsis siliques. I am facing difficulties in extracting RNA from the limited amount of sample (approx. 30-50 mg of...

02 December 2017 3,175 3 View

How much of the genomic DNA and probe should I use with Alkphos labelling and detection kit to detect transgene copy number in arabidopsis?

I am trying to detect the transgene copy number in Arabidopsis thaliana ecotype col 0 with Alkphos labeling and detection kit. As per the manufacturer's instructions I am using 20ng probe per ml...

21 June 2014 8,451 2 View