Giacomo Pesci

6 Questions 1 Answers 0 Followers

Questions related from Giacomo Pesci

How to manipulate the transition from exponential to stationary phase in mammalian cell culture?

Is there a way to stop exponential growth and increase protein production, without using chemical inducers? What can happen if one increases/decreases the concentration of nutrients in the medium...

03 April 2020 9,680 0 View

How frequently do polymerases make mistakes?

I used the Gibson assembly to join together two PCR fragments (4400 bp +770 bp). After transformation of E. coli with the Gibson mix, I isolated DNA from single colonies and I performed a...

29 March 2020 1,477 3 View

Which are the most effective ways to promote oxidative metabolism during cell culture?

I am looking for chemicals that can be used in mammalian cell culture to enhance oxidative metabolism (TCA cycle). Can NAD+ precursors (NR, for example) be used?

18 March 2020 7,520 4 View

Purification of fusion protein with Protein L affinity column?

I am studying a single-chain variable fragment (scFv) linked to a luciferase (Nanoluc) at the C-terminus (N-scFv-linker-Nluc-C). The naked scFv (not linked to Nluc) can be easily purified using...

23 February 2020 9,209 2 View

Purification of fusion protein with Protein L affinity column?

I am studying a single-chain variable fragment (scFv) linked to a luciferase (Nanoluc) at the C-terminus (N-scFv-linker-Nluc-C). The naked scFv (not linked to Nluc) can be easily purified using...

02 February 2020 6,285 2 View

Why do my minipreps look like this?

I did several minipreps (different plasmids extracted from different cell lines) and when I run them on agarose gel 0.8% they don't look good (smear at high MW and it looks always like that). The...

06 June 2019 9,915 5 View