VirusEpifluorescenceProtocolFinal-2

VirusEpifluorescenceProtocolFinal-2 - Bi1x, Spring 2010...

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Bi1x, Spring 2010 Virus Epifluorescence Protocol In this experiment your group will be imaging viruses from pond samples using SYBR Gold. The samples are stained and then run through a 0.02 micron filter which is small enough to catch viruses. The filters will then be loaded onto a slide and imaged under the fluorescence microscope. Your group will prepare all four of the conditions described below. Each person should complete the entire process at least once. Conditions 1. Unmodified pond sample 2. Pond sample prefiltered a. Get a syringe and a .2 micron filter b. Remove the plunger, screw on the filter c. Add 2ml of sample and force it slowly and gently through the filter into a new labeled tube 3. Control viruses a. Get an aliquot of lambda phage at known concentration from the TAs 4. Milli-Q water filtered through a 0.02 micron filter Staining Protocol 1. Add 2ml of your sample into a 15ml falcon tube 2. We will have diluted the stock 10,000X SYBR Gold solution to 100X (into 100 uL of water that was prefiltered with a 0.02 micron filter – supplied by the TAs)
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VirusEpifluorescenceProtocolFinal-2 - Bi1x, Spring 2010...

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