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==Set up MDA reactions== *Perform serial dilution of GM20431 gDNA to 30pg/ul. *Perform UV treatment of ALS and NS buffer 10ul/tube for 5 min; *Prepare 2X SYBR Green I solution from the 50X stock solution; *Mix 1ul template with 1.5ul ALS buffer, RT for 3min; *Transfer to cool block; *Add 1.5ul NS buffer; *Add 16ul Master mix: x13 x9 Tre(-) Tre(+) Trehalose(1.2M) 0ul 5.0ul 10x RepliPhi phi-29 buffer 2.0ul 2.0ul 1mM primer 1.0ul 1.0ul 25mM dNTP 0.8ul 0.8ul 2X SYBR Green I 1.0ul 1.0ul H2O 9.7ul 4.7ul RepliPhi Phi-29 (100U/ul) 1.0ul 1.0ul *Incubate at 30C for 10h on Opticon4, take one fluorescent read every 6 minutes. [[Image:2009-02-12-MDA-N6.png|300px]] [[Image:2009-02-12-MDA-LN6.png|300px]] [[Image:2009-02-12-MDA-LN6-Tre.png|300px]] [[Image:2009-02-12-MDA-N9-Tre.png|300px]] [[Image:2009-02-12-MDA-LN6-37C.png|300px]]
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