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Human cell lysate MDA test 3
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===Lymphocytes pre-processing=== *Take cultured lymphocytes suspension from one T25 flask **Aliquot 100 uL suspension for cell counting using the cell counter (Beckman) **Withdraw the desired cell amount (2 x 10^5 cells) and wash using UV-treated 1XPBS twice (Spin at 1000rpm for 3min) **Resuspend in 1 mL UV-treated PBS **Make dilution to 10 cell/uL using UV-treated PBS: Dilution Factor Start 1/10 1/40 1/400 ---------------------------------------------------------- BAC mixture 100 uL 100 uL 10 uL 10 uL PBS 900 uL 30 uL 90 uL ---------------------------------------------------------- Concentration 200/uL 20/uL 5/uL 0.5/uL Mix by repeat pipetting using p200 pipettor in each step *Realtime PCR program set up (Number of reactions) **cell lysate: 0.5-cell x 24 **Positive controls: 6 No-template control: 2 **Total reaction: 32 x 20ul
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