Top 10 cells

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Felixke
Felixke's picture
Top 10 cells

Hallo,
a newbe question: what are TOP10 cells.
I saw this in an article I needed, but I have no idea what they mean with Top10 cells.
Any ideas?
I think it has something to do with some sort of genetic factor?
 

heehawmcduff
heehawmcduff's picture
I think this is pretty
Felixke
Felixke's picture
I allready saw that, but

I allready saw that, but didnt really made me wiser.
They allready start with top 10 cells:
Preparing seed stocks

  • Streak TOP10 cells on an SOB plate and grow for single colonies at 23°C
    • room temperature works well

So they allready strak top 10 cells... but still makes me wonder: what are top 10 cells?
 

kjosephs
kjosephs's picture
Top10 cells are a strain of E

Top10 cells are a strain of E. coli used for cloning and plasmid preparation.  They have a high transformation efficiency and have the genes for recA (a recombinase) and endA (a DNAse that lowers plasmid yeild) knocked out.  I think "TOP10" is the brand name for the product sold by Invitrogen and it is very similar to other cloning strains like DH5alpha.

Felixke
Felixke's picture
Hallo kjosephs,

Hallo kjosephs,
Thats very intersting.
Do you have some sort of reference of this? Some article?
I have been looking but couldnt really find anything on it.
cheers

Fraser Moss
Fraser Moss's picture
The TOP10 E.Coli strain  are

The TOP10 E.Coli strain  are commercially available from invitrogen.
their genotype is :
F- mcrA Delta(mrr-hsdRMS-mcrBC) φ80lacZDeltaM15 DeltalacX74 recA1 araD139 Delta(ara-
leu)7697 galU galK rpsL (StrR) endA1 nupG
 
You can read a manual here
http://tools.invitrogen.com/content/sfs/manuals/oneshottop10_man.pdf
 
BEWARE the different substrains though like TOP10 F'
which posesses the F' episome lacking in the TOP10's described above
their genotype: F´{lacIq Tn10 (TetR)} mcrA Delta(mrr-hsdRMS-mcrBC) Φ80lacZ?M15 DeltalacX74 recA1
araD139 Delta(ara-leu)7697 galU galK rpsL endA1 nupG
 
tools.invitrogen.com/content/sfs/manuals/oneshottop10f_man.pdf

Felixke
Felixke's picture
Those are nice links, thanks

Those are nice links, thanks a lot frasermoss.
I suppose those top 10 cells are the same as the top 10 chemical competent cells, since they use both on the invitrogen website.
 
 

Fraser Moss
Fraser Moss's picture
yes, they just prepare them

yes, they just prepare them differently depending on the application for which they will be used

Felixke
Felixke's picture
oh, they are different then.

oh, they are different then.
They prepare the cells like you want them? Depening for what you need them, they can adjust the cell line then and make them chemicaly competent like you wish?
thanks for the info.
 

Fraser Moss
Fraser Moss's picture
the number of cells and the

the number of cells and the buffer in which they are frozen ready for use are different depending in whether you want chemically or electrocompetant cells.
The genotype of the TOP10 cells will be the same

Felixke
Felixke's picture
Aha, I see.

Aha, I see.
Thanks a lot frasermoss!! I understand it now.
Cheers

varsha
varsha's picture
I am moving this post to

I am moving this post to "Cloning" forum.
Varsha

cdurre
cdurre's picture
I learned something new about

I learned something new about detergent from the link provided about competent cells. Informative and interesting.
http://openwetware.org/wiki/TOP10_chemically_competent_cells 
 

dhivergi
dhivergi's picture
IS THIS E ecoli TOP10 doesnt

IS THIS E ecoli TOP10 doesnt have plasmid inside their chromosome