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====CoRE Fragmentation protocol==== *Follow [[EricChu:MDA/CoRE fragmentation protocol for microfluidic device|Eric's protocol]] * Prepared reagents dilution and kept on ice # 3.3x exo- Klenow buffer (from epicentre): 30ul 10x buffer + 90ul water # diluted UDG: 1ul UDG (5000U/ml) + 128ul water # diluted endonuclease IV: 1ul endoIV (10,000U/ml) + 128ul water # diluted exo- Klenow: 1ul exo- Klenow + 50ul water # diluted dNTP (from epicentre): 1ul 25mM dNTP + 25ul water *In strip PCR tubes, added # 1ul 3.3x exo- Klenow buffer # 1ul diluted UDG # 1ul diluted endonuclease # 1ul sample from MDA device * 37C for 2 hours and 65C for 15min * Then added to PCR tubes # 1ul diluted exo- Klenow # 1ul diluted dNTP * 37C for 1 hour and 75C for 15min
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