Sergei Smirnov

7 Questions 5 Answers 0 Followers

Questions related from Sergei Smirnov

Could you please suggest why Hek293 cells lose fluorescent protein expression upon bsd selection after lentiviral transduction?

Could you please advise me what could be the reason of loss of fluorescent protein expression? As a vector I take a lentiviral plasmid pUltra-hot(247629) in which the fluorescent protein mCherry...

28 November 2024 1,440 0 View

Can anyone suggest why the D283 medulloblastoma cell line did not attach after thawing?

Can anyone suggest why the D283 medulloblastoma cell line did not attach after thawing? What changes can be made to the dish surface or culture medium to solve this?

01 December 2023 3,373 2 View

Additional band near upper marker during NGS library preparation from FFPE?

Good day, could anybody suggest what could be the reason of a additional big band need upper marker? If we exclude contamination. I am using Agilent sure select XT protocol for targeted DNA seq of...

05 April 2022 7,035 0 View

What is normal percentage of successful lambda phage infection(lytic) ?

Could you please tell, what percentage of cells are usually successfully infected(lytic infection) after first round of infection in bulk culture(after 50 minutes). When you work with DH5alpha...

09 March 2020 1,957 0 View

Why in liquid culture I have lower protection level of restriction modification system from lambda phage then on the plates?

I am infecting E.Coli(DH5alpha) with lambda EYFP with amber mutation that doesn't allow to lyse cell, but it can multiply inside. (30min on ice and 1,5 hour at 37,MOI=1). When I calculate the...

04 January 2020 6,294 3 View

What could be the reason that the level of protection of RM system in liquid culture is 10 times less than on the plates?

I am infecting E.Coli(DH5alpha) with lambda EYFP(30min on ice and 1,5 hour at 37,MOI=1). When I calculate the number of infected cells using both microscopy ant FACS I see that in samples with RM...

16 November 2019 4,127 3 View

What strain to use in cloning with araBAD promoter and gene of interest(1000bp) that is complementary to the region on the same plasmid?

I am trying to insert into PBR322 araBAD promoter with my gene of interest(it should be inducible). Could you please tell wether I could use DH5alpha strain as I was told it lacks arabinose...

29 January 2019 3,556 4 View