Megha Achar

10 Questions 41 Answers 0 Followers

Questions related from Megha Achar

After purifying protein of interest their is non-specific multiple bands above and below the protein of interest band. Can DEAE method be used?

I am trying to purify a protein and after purification the expression of protein of interest is pretty good but their are non-specific bands below and above the protein of interest. Can anyone...

09 September 2015 3,029 12 View

I had done PCR standardization to amplify my gene of interest of 1.5kb from cDNA. When I repeated the same protocol, no amplification was seen. Why?

I had done PCR standardization using cDNA as template. I did too many pcrs and finally standardized to amplify the gene of interest of 1.5kb from cDNA. I amplified 2 different genes but of same...

11 November 2014 2,990 8 View

I did cloning for phusion protein. I did westerns and getting a band of interest along with a band with low molecular weight, why?

I had cloned my gene of interest in to a vector, which has fluorescent protein tag. I did restriction digestion and did insert release of 1.4kb and fragment release having both gene of interest...

07 July 2014 6,242 8 View

Why am I not able to get a clone of gene tagged with GFP?

As previously discussed, I redesigned the primers. I tried triple ligation, it did not work. So, the gene of interest was restriction digested from the vector containing gene and was amplified...

01 January 2014 675 9 View

I am not getting a desired amplification for new set of gene specific primers designed?

Forward primer: 79.6 degrees melting temperature and Reverse primer: 81.1 degrees. Reverse primer has strong secondary structure and no dimer. I tried pcrs setting with high denaturation temp and...

11 November 2013 2,227 16 View

How to write scientific paper related to clinical psychology in neurological disorders?

I have collected the data and analysed. But writing discussion is little difficult, as I am looking for pathology in brain for psychological symptoms. Please suggest me as to how the discussion...

09 September 2013 9,607 5 View

PCR reaction is not working and not showing any amplifications, while positive control shows. I changed buffer, Mgcl2, dNTPs mix, water. Why is this?

I have been doing genotyping for a mouse model. I isolate the genomic DNA from Alzheimer's mice tails. I have done many PCRs, but this batch of isolated genomic DNA, has good concentration and...

09 September 2013 9,551 49 View

How to maintain a cell line N2a?

I see different people using different ways to maintain the cell culture. Can anyone please explain the basic steps involved in cell culture maintain, passaging etc. As to how much cells should be...

07 July 2013 594 2 View

Can I use a combination of acids like Hcl and H2so4 / Hno3 in adjusting the pH of Tris buffer saline?

Tris buffer saline of pH-7.4. As in case of emergency like one of the above mentioned acid gets over while adjusting the pH, than rest of the pH could be adjusted using other alternative acid. Is...

07 July 2013 7,725 2 View

In PCR, why do the salts differ for particular polymerase enzymes like Mgcl2 for Taq and MgSo4 for Kod polymerase?

However both salts have Mg ions.

06 June 2013 7,223 2 View