User:Karmella Haynes/Notebook/Polycomb project/2010/07/23

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07/23/10

 * &#x2713; RNA prep: for Nanostring assay (cDNA synth. on Monday)

RNA prep > Use Qiagen QIAshredder and RNeasy kit (no BME added to lysis buffer) > Elute & re-elute with 30 μL RNase-free H2O

--> Store at -80°C *Note: use some for cDNA for RT-PCR confirmation of Nanostring results

7/26/10 > oligo(dT) Primer annealing

> cDNA synthesis mix --> 10 reactions total

--> Add 10 μL mix to each annealing rxn. --> 50°C/ 50 min., 80°C/ 5 min., ice --> Add 0.8 μL RNase H, 37°C/ 20 min. --> Store at -20°C


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