Matt:LabNotes/2014-2-10: Difference between revisions

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===Quantify Capture===
===Quantify ppMALAT1 in vitro Capture from [[Matt:LabNotes/2014-2-6 | 2/6/2014]]===
*68.8 ug/ml
 
*75.3 ug/ml
'''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 about -> 2.69 uM

Revision as of 23:12, 10 February 2014

Quantify ppMALAT1 in vitro Capture from 2/6/2014

Samples

  • 8 PCR tubes of 35ul from 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 about -> 2.69 uM