Priyanka G Bhosale

10 Questions 21 Answers 0 Followers

Questions related from Priyanka G Bhosale

How can I remove a non specific band at a particular molecular weight in western blotting?

The protein of my interest is of 69 k Da, but i observe a nonspecific band at 55 Kda along with the specific band, even after increasing blocking i see this band, it also stains ladder at same...

09 September 2014 9,559 4 View

Can anyone suggest best transfection method (non viral) for SCC29B (buccal mucosa cell line)?

I want to down-regulate gene of interest in SCC cells but this cells are hard to transfect. I have tried using Xtreme gene as well but got 10-15% transfection.

08 August 2014 5,420 3 View

Can anyone suggest me sequencing condition for a shRNA-plasmid forming strong hairpin loop?

sh-RNA for a gene is cloned in pLKO1.puro vector and it foams a strong hairpin loop, due to which sequencing reaction terminates abruptly. Please suggest me what can be done in this situation.

02 February 2014 5,411 2 View

Protocol for detection of Phospho EGFR by IHC

I am standardizing IHC for pEGFR detection using vector ABC kit, and tried different retrial conditions and antibody concentrations, but I'm unable to get staining. Can anyone suggest any specific...

07 July 2013 175 2 View

Has anyone tried - AllPrep DNA/RNA/miRNA Universal Kit from QIAGEN (Cat no 80224)

This column extracts DNA and total RNA (mRNA +miRNA), so had a query that whether miRNA so obtained can be used for miRNA profiling.

04 April 2013 3,792 0 View

Which is best PCR primer for HPV (integrated or episomal) detection in oral tissues.

I use Nested PCR with MY09/11 followed by GP5+/6+ primers, but if anyone has a better set than these please let me know as prevalence of HPV in gingivobuccal cancer is very low, so I don't want to...

03 March 2013 9,523 5 View

Non specific band in PCR for HPV detection, what can be done about it?

I use PCR to detect presence of HPV in my tissue samples, In positive sample i get single specific band (150 bp) but those samples which are negative i get higher mol wt non specific multiple...

11 November 2012 5,953 29 View

How to extract DNA and RNA from epithelium?

Normally I use QIAGEN kit for extractions from tumor tissues, but now I am planning to do it from premalignant lesions where I need only epithelial cells. I tried taking serial cryo sections and...

10 October 2012 8,192 21 View

What is the best protocol to treat culture with 5-aza-2'-deoxycytidine to block methylation?

I am planning to block methylation in my culture and check its effect on expression of particular gene, so how should i treat my culture with aza cytidine? i.e should I give continous dose or at...

10 October 2012 1,473 18 View

Is it possible to determine the reason why one of my genes of interest is down-regulated, eg promoter methylation or something else?

In my study on human tumors, samples showed down regulation of one of my genes of interest. I am interested to know whether its because of promoter methylation or transcriptional inhibition. How...

10 October 2012 2,902 9 View