16 Questions 58 Answers 0 Followers
Questions related from Dr. Sankareswaran Senthil Kumar
Dear Research friends, i have one doubt about FBS (fetal bovine serum) storage condition?.. Once you are all heat inactivated your serum and then aliquote 50 ml each to store at -20C . Why? Once...
11 November 2014 656 8 View
In our lab, Sanyo-MCO-175 water jacket Co2 incubator was used for cell culture maintenance. Past 3 month we were faced contamination problem in our cell lines (A549 and MCF7). So I was checked all...
08 August 2014 7,542 8 View
I have doubt about our lab south american FBS because it will be expired march 2015. I have a doubt about FBS also. In my A549 culture this one gave rad and spiral shape contamination. So can you...
07 July 2014 1,803 19 View
I want to use agarose as a matrix for this assay on behalf of collagen or fibrin. Is it possible? If I use agarose as a matrix does that mean I can add any specific growth factor?
12 December 2013 2,324 3 View
I used GENET BIO (ExPrime Tag Premix (2x)) for my PCR rxn. It contains: DNA Polymerase 1 Unit/10 microliter, 20 mM Tris-HCL, 80 mM KCl, 4mM Mgcl2, enzyme stabilizer, sediment loading dye, pH- 9.0...
10 October 2013 9,457 4 View
I didn't get my clone (pcDNA3.1+insert+GFP). I cloned GFP successfully but my promoter cloning is very difficult . I replaced CMV promoter with my promoter (2 different promoter (1).930 BP and...
08 August 2013 4,523 1 View
i have one more doubts.. we were maintained A549, MCF7, INS cell lines if i should maintain upto 50 passage or more. 1). Normally How many passage cells are used MTT or some other assay? 2). If we...
07 July 2013 9,238 13 View
I was used different concentrations (100, 50, 25, 12.5, 6.25, 3.125 µg/ml) of particulate matter (PM10) treated with A549 cell line for 24 hours, 48 hours. I was completed 24 hours reading at 570...
07 July 2013 8,746 13 View
I analyzed particulate matter (PM) sample from Selam industrial area, Tamil Nadu, India. I added my particulate matter in various concentration to the A549 cells. I used 96 well plates. After 16...
07 July 2013 7,530 0 View
1). Wound healing 2). Cell proliferation 3). Apoptosis 4). Etc... I have A549, MCF7, HepG2, THP1 . So please suggest to me, which passage number is suitable for above assays? My work :...
07 July 2013 5,472 12 View
I have pcDNA3.1+ Insert + GFP vector. I transfected this vector into A549 cell line. I got minimum GFP expression after 48 hours (only 5 to 10 cells give GFP fluorescence) and I trypsinized that...
07 July 2013 2,146 12 View
I am using Sac I (NEB BIO labs) RO 156S 2000 units - (20,000 U/ml)). How many microliters will need for 20 Microliter Restriction Reaction? (2 microgram of DNA). Sac Data sheet didnt give proper...
06 June 2013 8,999 11 View
I have 20 BP forward primer and 24 BP Reverse Primer and also my product size oligos (63 bp & 79bp). I used my primers to amplify my products. Finally I got my exact size of the products in gel...
06 June 2013 4,651 30 View
We are facing a lot of contamination problems.
05 May 2012 3,097 5 View
I have THP1 culture but it was contaminated. I have used pen-strep (Sigma) and also tetracycline antibiotic solution but still it didn't kill the contaminant (Brownian movement). Can anyone offer...
04 April 2012 7,987 15 View
In my observation cell morphology is totally changed.
03 March 2012 5,704 2 View
