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===Prep=== * Tn5059 was supplied at 25X, with transposons A and B in separate tubes. We are assuming that a 50/50 mix of A/B is 25X. We have both control transposons and T7 promoter transposons for Brandon's IVT project * Follow the latest Nextera DNA sample prep user guide for tagmentation conditions and PCR. To avoid purification after tagmentation, use Qiagen Protease to inactivate the Tn5 * Dilute 25X Tn5 A/B mix to 5X in UV treated TE buffer with 50% glycerol before use. 1X Tn5 reaction in 5 ul has shown good fragment distribution for 6-8 ng of purified DNA (MDA product), but appears to over-fragment smaller template inputs ** This is similar to older Nextera * Volumes below are per reaction. Make master mix with ~10% excess for multiple reactions * Include both positive control (purified DNA with known concentration), NTC (No Template Control), and NEC (No Enzyme Control) * UV treat tubes, buffer, and H2O before use
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