2 replies to this topic

[culturenewbie]

Hi,
I've just been given the task of setting up a reporter gene experiment with mouse kidney cells.  
I've looked up methods etc. on the likes of the Promega web site but haven't got a clue where to go for mouse kidney cell lines. Also have a small problem of not having any equipment to grow the cell lines when I get them and very limited budget to get what I need. Can someone please give me an idea of the minimum equipment requirements to culture cells, where I could look to find such cell lines and any helpful hints on how to set about it all? My only experience of cell work is at least 20 years old so I'm starting from scratch here. Any help gratefully appreciated
  
Fiona (culturenewbie)

[postdoc2130]

First, you need cells and proper media.
ATCC (American Type Culture Center) is the place to find your prefer cell lines and appropriate media for carry the cells.

Then, you need cell tranefection kit to put the reporter gene in. My lab has been using Epoch Biolabs' GenCarrier-1 or GenCarrier-2 (depend on with serum or serum free condition). These reagents has been successfully worked on many established cell lines and many primary cells. In my hand, it work the same as lipofectamine2000 but only half price ($180 for 250 Tfx).

Next, you need a reporter assay to monitor reporter gene expression. All depend on which reporter you prefer (usually, luciferase, CAT, SEAP, GFP or LacZ).

You also need a culture hood, CO2 incubator and centrifuge for your cell prep. Trypsin, PBS (w/o Ca, Mg), FBS....Cryopreservation - small tank....

[BryanW]

you may also need petri dishes (35-, 60, 100-mm) and 6-, 12-, 24, 48, 96-well plates, serological pipettes (2-, 5-, 10-, 25-ml) for your experiments.
Fisher Scientific always has cheaper price on plastic wares.

With your tight budget (like our lab), I believe GenCarrier-1 is your best choice you can find (our lab has been using GenCarrier-1 on pancreatic PANC-1 and primary pancreatic islet cultures. It works perfectly!!!). In addition, for transfection, you need extra-pure DNA. We use GenCatch maxi kit also from Epoch Biolabs (epochbiolabs.com). it's cheaper.