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===Purify UHRR of DNA=== *Use Zymo DNA-Free RNA Kit *Sample: 4ul of UHRR (1.3 ug/ul) ====DNase I Digest==== {| {{table}} | align="center" style="background:#f0f0f0;"|'''Components''' | align="center" style="background:#f0f0f0;"|'''Volume''' |- | UHRR (1.3ug/ul)||4 |- | 10X DNase I Buffer||5 |- | DNase I||2 |- | H2O||39 |- | Total||50 |} *Mix and incubate at 37C for 30min ====Purify==== *Add 100ul RNA Binding Buffer and mix *Add 150ul 100% EtOH and mix *Transfer to Zymo-Spin IC Column in Collection Tube **Centrifuge at 13,000rcf for 1min and discard flow through *Add 400ul RNA Prep Buffer **Centrifuge at 13,000rcf for 1min and discard flow through *Add 800ul RNA Wash Buffer **Centrifuge at 13,000rcf for 30sec and discard flow through *Add 400ul RNA Wash Buffer **Centrifuge at 13,000rcf for 30sec and discard flow through *Centrifuge at 13,000rcf for 2min and discard flow through *Transfer column to 1.5ul tube and add 8ul H2O and let stand for 1min **Centrifuge at 10,000rcf for 30sec *Store at -20C
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