Biomod/2014/OhioMOD/experimentnotes

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Experiment Notes


Table of Contents

1. Cellular Uptake Experiment (DNA Extraction)v1
Data 2
Data 3



1.Cellular Uptake Experiment (DNA Extraction) v1


07/29/2014


Materials:


  • 18 mM Branch folded 7/16
  • OSU CLL cells
  • Qiagen DNeasy Blood and Tissue Kit
    Procedure:
  • The concentration of Branch was determined using Nanodrop
      The concentration was 1.5 nM.
  • 200 uL of OSU CLL cells were washed and incubated in cell media.
  • 50 uL of structure was added to the cells, and solution was plated and incubated overnight.
      Final volume: 250 uL
      Final concentration of structures: 0.3 nM



    07/30/2014

  • The cells were washed once in PBS, then the total DNA was extracted from the cells according to the Total DNA extraction protocol.
  • A gel was run with the extracted DNA, purified branch structures, 7249 scaffold, and ladder.
    Results: <figure>
    <img src="http://openwetware.org/images/e/e5/Gel_verification.jpg"height="300" width="300"/> <figcaption>Fig: Gel verification of DNA extraction. From left to right: ladder, scaffold, purified branch structure, DNA extract.</figcaption> </figure>


    As the gel shows, the DNA extract had no band that corresponded with the branch purified structure.

    Discussion:
    Since there was no band on the gel corresponding with the folded branch structures, there is no way to confirm whether the structures were uptaken or not and whether the DNA extraction was able to extract the DNA origami structures or not. If structures were uptaken, then the concentration of structures might have been too dilute to register on the gel, since the initial concentration in solution was only 0.3 nM.


    For future experiments, the cells should be incubated in a higher concentration of structures. In addition, TEM imaging can be used to determine whether there are any structures present in the DNA extracted from the cell.





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