Difference between revisions of "Part:BBa K1777000:Design"
 
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===Design Notes===
 
===Design Notes===
 +
miR-21 is overexpressed in Hela cells and U87 cells according to previous report.<br>
 
We use primer annealing to synthesize the mir-21 binding domain with Xho I cohesive end and Pme I blunt end. Cut the psiCHECK2 vector with the same two enzymes, then use T4 DNA ligase to construct the plasmid.<br>
 
We use primer annealing to synthesize the mir-21 binding domain with Xho I cohesive end and Pme I blunt end. Cut the psiCHECK2 vector with the same two enzymes, then use T4 DNA ligase to construct the plasmid.<br>
 
In order to make sure of this reporter's efficiency, we synthesize two binding sites which are combined together.
 
In order to make sure of this reporter's efficiency, we synthesize two binding sites which are combined together.
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siCHECKTM Vectors INSTRUCTIONS FOR USE OF PRODUCTS C8011 AND C8021.(Promega)<br>
 
siCHECKTM Vectors INSTRUCTIONS FOR USE OF PRODUCTS C8011 AND C8021.(Promega)<br>
 
Dual-Luciferase® Reporter Assay System Instructions for use of Products E1910 and E1960.(Progema)
 
Dual-Luciferase® Reporter Assay System Instructions for use of Products E1910 and E1960.(Progema)
 +
Jennifer A. Chan,1,3 Anna M. Krichevsky,2 and Kenneth S. Kosik4,et al.MicroRNA-21 Is an Antiapoptotic Factor in Human Glioblastoma Cells.Cancer Res 2005; 65: (14). July 15, 2005

Latest revision as of 10:09, 20 September 2015

two mir-21 binding sites


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

miR-21 is overexpressed in Hela cells and U87 cells according to previous report.
We use primer annealing to synthesize the mir-21 binding domain with Xho I cohesive end and Pme I blunt end. Cut the psiCHECK2 vector with the same two enzymes, then use T4 DNA ligase to construct the plasmid.
In order to make sure of this reporter's efficiency, we synthesize two binding sites which are combined together.

Source

Renilla Luciferase and Firefly Luciferase,as the control insert of this part both come from psiCHECK-2 Vector.

References

siCHECKTM Vectors INSTRUCTIONS FOR USE OF PRODUCTS C8011 AND C8021.(Promega)
Dual-Luciferase® Reporter Assay System Instructions for use of Products E1910 and E1960.(Progema) Jennifer A. Chan,1,3 Anna M. Krichevsky,2 and Kenneth S. Kosik4,et al.MicroRNA-21 Is an Antiapoptotic Factor in Human Glioblastoma Cells.Cancer Res 2005; 65: (14). July 15, 2005