Nikhil Shah

10 Questions 15 Answers 0 Followers

Questions related from Nikhil Shah

What amount of construct DNA should I use in the nucleofection protocol for CRISPR-Cas9 gRNA?

I want to perform the nucleofection of CRISPR/Cas9(WT) gRNA construct in beta cell line. 1. How much construct DNA shall I use for the nucleofection. Technique- Amaxa Nucleofector Kit- T or Kit-...

05 May 2015 2,169 5 View

How do I solve my problem in generating Bacmid?

I am generating bacmid in DH10 Bac cells. When I transformed the DH10 Bac cells, I got blue white colonies after 72 hr, but the colonies were very few just 4-5 white colonies. White colonies were...

05 May 2015 8,732 4 View

What Bioinformatics tool can I use for PCR efficiency check ?

I am looking for tool to evaluate the PCR efficiency for any template/gene. Especially to get an idea about the nature/complexity of gene stretch e.g. Any stable secondary structures, polymerase...

04 April 2015 2,347 4 View

How can I digest only one RE site of vector if having two sites of the same REs?

I am using pIRES bicistronic vector for cloning, where it contains two sites of Sma-I in MCS-B region. I have designed cloning primers with sma-I forward and Sal-I reverse. Since another sma-I...

03 March 2015 555 10 View

Concept of Bed volume in protein purification ?

Please explain the concept of bed volume in protein purification and how to calculate it ? E.g. I am using glass Econo Column(1.0X10cm)Biorad#737-4011 and using Ni-NTA slurry of 0.5 ml or 1mL....

02 February 2015 6,525 1 View

How can I improve the purity and quantity of his-tagged protein?

I am purifying one of the tyrosine kinase protein expressed as his-tagged in SF-9 cells, however I am facing problems in Ni-NTA protein purification. The details of the procedure is as...

12 December 2014 1,509 16 View

What is the ideal lysis buffer composition for colon cells like FHC and CCD-18Co?

I tried buffer: 1%NP40, 20mM Tris, 137mM Nacl, 10%glycerol, 2mM EDTA, 1mM activated Na-Orthovanadate, Aprotinin 10ug/ml,leupeptin 10ug/ml. Incubation with lysis buffer for 30min but still I see...

04 April 2014 8,338 4 View

How can I perform a search in order to find out the promoter region of any gene?

eg. if I want to search the promoter for IGF1 gene.

03 March 2014 4,246 4 View

Can anyone guide me on how to search for the promoter region of any gene??

Molecular biology

03 March 2014 5,374 5 View

Has anybody worked on the detection of IGF1 protein in cell culture media?

Assay development

08 August 2013 6,143 2 View