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If you do have to use an auxotroph, be very careful with the supplements you add. For example, you can't use adenine sulfate with sulfur limitations. You want
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Block with RPMI (1X) + 1X ELISA/ELISPOT buffer at RT for 2 hrs Remove media and add 20 ml of HBSS (serum free); spin cells for 5 min at 1400 rpm.