Marcelino Telechea Fernández

4 Questions 7 Answers 0 Followers

Questions related from Marcelino Telechea Fernández

How to prevent DNA contamination in a Yeast Maxiprep?

We performed a transformation-associated recombination protocol in yeast of lot of DNA fragments and the vector to get a YAC of aprox 35KB. We first checked the proper assembly of the fragments by...

18 December 2020 8,681 0 View

How to overcome to a big differences between biological replicates of rRNA expression in qPCR?

I use b-actin as my housekeeping and I don´t have that problem, and the technical replicates (pippeting) of the 47s rRNA work fine as well, so...I am about to try with the third pair of primers,...

24 May 2017 2,890 2 View

Is there any specific rule to design the dissociation stage (melting curve) at the end of running a qpcr with sybr green?

I mean, I am not able to find if there´s something like a general protocol or the time and the temperatures change according to your conditions? I guess the latter is the correct option but...how??

01 March 2016 8,915 2 View

Does anyone know why the protease inhibitor leupeptine is used in calpain casein zymograms when it is a cystein protease inhibitor too?

I am checking several protocols and all of them use leupeptine in their buffers, and I don´t understand how is possible to accurately assess the calpain activity if you are inhibiting it...thanks

07 January 2016 2,879 5 View