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[DESCRIPTION]
Purpose: to measure the infective titer (number of infective elements per mL) of from 1-5 viral samples in a quick and efficient manner.

[GUIDELINES]
<strong>(Modified from R. Fu, K. Frois-Moniz &amp; M. Sullivan protocol)<br /><br />Materials: (assuming 2 replicates per viral sample)<br /></strong><br />&bull; Two sterile, flat-bottom, 96-well plates for up to 5 viral samples with a capacity for &gt;300 &micro;L<br />&bull; 30 mL Pro99 or SN medium for each dilution plate plus 30 mL for each sample test plate<br />&bull; 7 mL late-exponential phase host cells (~3x10<sup>7</sup>/mL)<br />&bull; 60 &micro;L each virus sample<br />&bull; Multi-channel pipettor with 12x300 &micro;L and 8x1500 &micro;L heads and a 20-100 &micro;L pipette<br />&bull; Sterile tips for pipettors<br />&bull; Sterile plastic troughs for cells and growth medium for use with multi-channel pipettors<br />&bull; Parafilm for sealing lids to prevent evaporation<br /><br /><strong>Labeling example for step 1</strong><br />
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<p><em><u>Virus Sample A</u></em></p>
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<p><em><u>Virus Sample B</u></em></p>
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<p><em><u>Virus Sample C</u></em></p>
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<p><em><u>Virus Sample D</u></em></p>
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<p><em><u>Virus Sample E</u></em></p>
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<p>1</p>
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<p><em>-2 </em>&nbsp;&nbsp;A</p>
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<p><em>-3</em>&nbsp;&nbsp; B</p>
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<p><em>-4</em>&nbsp;&nbsp; C</p>
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<p><em>-5</em>&nbsp;&nbsp; D</p>
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<p><em>-6</em>&nbsp;&nbsp; E</p>
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<p><em>-7</em>&nbsp;&nbsp; F</p>
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<p><em>0</em>&nbsp; G</p>
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<p><em>-1 </em>&nbsp;H</p>
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<br /><br /><strong>Calculations:</strong> for calculation purposes, each well contains 30 &micro;L of cells mixed with 20 &micro;L phage, diluted with 200 &micro;L medium for a total of 250 &micro;L.<br /><br /><strong>Note:</strong> once you have prepared the phage dilutions, the rest of the procedure should be carried out as quickly as possible to minimize (a) possible viral adhesion to the plates and (b) the amount of time that the host cells spend out of the incubator.

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