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===Quantify ppMALAT1 in vitro Capture from [[Matt:LabNotes/2014-2-6 | 2/6/2014]]=== '''Samples''' *8 PCR tubes of 35ul from [[Matt:LabNotes/2014-2-6 | 2/6/2014]] **Actual volume ~30ul due to evaporation ====First trial of quantifying==== '''Procedure''' *Took the first and last tube *Brought each sample to 50ul with 20ul H2O *Used Zymo ssDNA/RNA Clean & Concentrate Kit following standard procedure *Eluted with 15ul H2O and quantified each with 1ul using Qubit ssDNA '''Results''' *Starting mass of linear ppMALAT1 in each tube: 3ul * 1434 ng/ul = 4.302 ug *Tube1: 68.8 ng/ul **15ul * 68.8 ng/ul = 1032 ng = 1.032 ug **'''Yield''' = 1.032/4.302 = 24% **1.032 ug * (umole/28019.4ug) = 0.0000368 umole = 36.83 pmole ** 36.83 pmole/15 ul = 2.46 pmole/ul = 2.46 uM *Tube2: 75.3 ng/ul **15ul * 75.3 ng/ul = 1129.5 ng = 1.1295 ug **'''Yield''' = 1.1295/4.302 = 26% **same calculation as above -> 2.69 uM ====Run Gel==== *Run denaturing TBU gel to check circularization **Lane 1: 0.5ul Low Mass ladder **Lane 3: 2ul 2uM ppMALAT1 linear ssDNA **Lane 4: 2ul 2uM ppMALAT1 linear ssDNA after Zymo ssDNA purification (from [[Matt:LabNotes/2014-2-5 | PosCtrl]]) **Lane 5: 2ul of circularized ppMALAT1 (from [[Matt:LabNotes/2014-2-5 | 1000ng capture]]) **Lane 6: 0.5ul of circularized ppMALAT1 from Tube 1 (68.8ng/ul) [[File:2014-02-10_ppMALAT1_Capture_Check.jpg]] ====Purifying the rest==== '''Procedure''' *Raised the volumes of the other 6 tubes to 50ul with H2O *Combined them into 3 tubes of 100ul *Used Zymo ssDNA/RNA Clean & Concentrate Kit *Eluted with 15ul each -> 45ul total
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