Jie:LabNotes/ASE/2009-6-10

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Revision as of 17:08, 10 June 2009 by >Jie deng (New page: ==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: Hybri...)
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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;