Iván Domènech Mercadé

6 Questions 5 Answers 0 Followers

Questions related from Iván Domènech Mercadé

Problems with BL21(DE3) pLysE bacteria transformation?

I am trying to transform the BL21(DE3) pLysE Chemically Competent Cells with my bacterial plasmid showed bellow which is Ampicilin resistent and expressing my genes of...

06 June 2018 3,496 7 View

Anyone has the faintest idea of why could be due to this strange and funny band shape on my agarose gel?

I performed a reverse PCR from a cDNA plasmid template in order to skip one exon of my gene of interest. Product size seems the correct one (5.7 Kb). But I don't know the reason why the bands on...

10 October 2017 2,872 11 View

Does anyone have experience splitting COS-1 Cells in 150 mm (10 cm) dish?

I would like to know which volumes You use to split a 100 % confluent 150 mm COS-1 cells dish in order to obtain an 80% confluent 150 mm dish 24 h after splting. Which volumes of tripsine do You...

07 July 2017 2,281 3 View

What power conditions (volts or mA) should I use in Western blotting for transferring a 15.6 kDa protein onto a 0.45 micron PVDF membrane?

The thing is that I am transfecting both protein isoforms separately in 10 cm dishes with COS-1 cells and FuGENE® HD Transfection Reagent. I am transfecting 3 µg of the each vector containing each...

03 March 2016 8,636 12 View

I am unable to get the band on Western blotting of the 15.6 kDa protein of interest isoform but I nicely detect its bigger 36.7 kDa isoform, why?

03 March 2016 9,838 6 View

In order to dectect LOH with an AB 3500 Genetic Analyzer, which size standard, kind of dye labelled primer and PCR product size is better to use?

I was considering about using a FAM-labelled Forward Primer, a GeneScan™500 ROX™ Size Standard or GeneScan™ 400HD ROX™ Size Standard, and a PCR fragment size of about 200-300 bp? It really makes...

03 March 2016 5,981 2 View