Mathies:Introduction to Cell Culture

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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 ly85@cornell.edu 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

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or instead, discuss this protocol.

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