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===Lambda Exo (Day 1)=== * Reagents used: 180 uL Lambda Exo (5 U/uL), 300 uL QIAEX II beads. * Set up time ~3 hr Reagent Volume dsDNA+water(~10ug) 125 uL Lambda Exo buffer,10X 15 uL Lambda Exo 10 uL 37C 1 hour -> 75C 10 min * I set up 3x150 uL reactions for A-D, and 6x150uL reactions for E. I expected <20ug after digestion, so after pooling 3 reactions together in a 2.0 mL tube, I purified with 50 uL QIAEX II beads and 3X volumes QX1 buffer. * Yield: {| {{table}} | align="center" style="background:#f0f0f0;"|'''Subset''' | align="center" style="background:#f0f0f0;"|'''Input(ug)''' | align="center" style="background:#f0f0f0;"|'''QIAEX II yield, Nanodrop (ng/uL)''' | align="center" style="background:#f0f0f0;"|'''QIAEX II volume (uL)''' | align="center" style="background:#f0f0f0;"|'''Total LambdaExo/QIAEXII yield (ug)''' | align="center" style="background:#f0f0f0;"|'''%yield''' |- | A||33.704||76.5||190||14.535||43% |- | B||28.248||62.5||190||11.875||42% |- | C||34.21||79.2||190||15.048||44% |- | D||35.42||81.5||190||15.485||44% |- | E||29.4679||67.1||190||12.749||43% |- | E||29.4679||69.1||190||13.129||45% |} * Check Lambda Exo efficiency by running PAGE TBE gel. ** LambdaExo digestion have good efficiency. ** QIAEX II can purify ssDNA [[File:460K_prep_May2015_A-E_LambdaExoDigestion.jpg | 300px]]
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