Mathies:Introduction to Cell Culture
Cell Culture Methods from Lina Yi
This section contains basic cell-handling techniques. Comment and modifications are always welcomed. Please feel free to contact me at firstname.lastname@example.org for any confusion I have made.
Before we start, here are some things that we need to prepare:
Important Reagents for protocols
Eppendorf tubes: All tubes should be autoclaved before experiments(autoclaved room: Lewis 108; tubes: Lewis 311 drawer “Eppendorf”)
Pipette and tips: All pipettes should be wiped with 70% ethanol before experiments and between cell transferring steps. We use pipette tips with cotton for cell handling (RT-10FG, ART 100E and ART 1000 Lewis 311 drawer “Eppendorf”)
Bacteria: E. coli strain K12 MG1655 (American Type Culture Collection, ATCC #700926) transformed with ampicillin resistance gene and nontoxigenic E. coli strain O157 NCTC 12900 (ATCC 700728); Lewis 311 freezer; stored 50µl aliquot of the grown bacteria solution per tube
Phosphate Buffered Saline (PBS): filtered through Millipore Steriflip (Catalog number: SCGP00525, placed in the cabinet of Lewis 310) in order to get rid of possible contaminations
- Eric Chu 22:15, 22 July 2009 (PDT):
or instead, discuss this protocol.