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Questions related from Laura Dionisi
Good morning, I am trying to produce a recombinant protein. Unfortunately I one of the two restriction enzymes that I use do not cut at the end of the MCS. After the digestion 15 nucleotides of...
18 July 2024 2,761 4 View
Hi I am trying to set up the reactions with my plasmid, insert and RE. Usually the standard final volume is 20 uL or 50 uL (it depends on the protocol provided by the manufacturer) with 1 ug of...
29 May 2024 1,040 3 View
I'm a doctoral student with early experiences with recombinant protein synthesis. Since I still have to start my experiment and I do not yet know the quantities of DNA and plasmid that I will...
20 May 2024 5,050 3 View
Hi My team and I are trying to design degenerated primers in order to sequence a viral genome. We do not know how many degeneration we can include in each primer. Is it possible to add up to 6/10...
23 February 2024 7,571 2 View
Hi, here in my lab I have a kit for RT-PCR with probes (for the real time quantification). Can I run my reaction without adding the probes, in order to obtain en “end point” product? I was...
01 February 2024 3,157 5 View
Hi, I have to perform a ligation reaction between a PCR fragment and a plasmid, in order to transform competent E. coli strain to amplify the plasmid. I will also make a gel to check the correct...
30 January 2024 9,237 7 View
Can anyone advise me on leader sequences to obtain recombinant proteins that are secreted by the cell culture? I should use mammalian cells for expression (from the Expi293 kit by ThermoFisher),...
09 January 2024 4,806 3 View
Hi! Has anyone worked with negative-sense RNA viruses? I need to express recombinant proteins using a vector, so it will be necessary to start with ssRNA(-) to obtain cDNA for selecting the gene...
02 January 2024 9,387 3 View
