Jie:LabNotes/ASE/2009-6-10
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Making ds-cDNA for Hues6/Hybrids/BJ
* Total RNA Samples: o A: Hues6-ES: 600ng/ul o B: Hues6-EB: 1ug/ul o C: Hybrid1-ES: 750ng/ul o D: Hybrid1-EB: 170ng/ul o E: Hybrid2-ES: 1.5ug/ul o F: Hybrid2-EB: 48ng/ul o G: BJ: 110ng/ul o H: Hybrid1-ES (diluted): 380ng/ul
Clean up and concentrate all RNAs with Zymo DNA-Free RNA kit
A B C D E F G RNA 8ul 5ul 6ul 30ul 3ul 80ul 20ul 10X DNase I buffer 5ul 5ul 5ul 5ul 5ul 10ul 5ul RNase-Free DNase I 2ul 2ul 2ul 2ul 2ul 4ul 2ul H2O 35ul 38ul 37ul 13ul 40ul 6ul 3ul 37C 10min
* Add 4 volumes of RNA binding buffer; * Transfer to Zymo-spin columns in collection tubes, spin @15k rpm for 30s; * Add 200ul RNA Wash buffer, spin @15k rpm for 30s; * Add 10ul DNase/RNase-free water, spin @15k rpm for 30s;