Difference between revisions of "Part:BBa K1777000:Design"

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<partinfo>BBa_K1777000 short</partinfo>
 
<partinfo>BBa_K1777000 short</partinfo>
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===Design Notes===
 
===Design Notes===
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We use overlap PCR to synthesize the mir-21 binding domain. In order to make sure of this reporter's efficiency, we synthesize two binding sites which are combined together.
+
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.
  
 
===Source===
 
===Source===
  
−
Renilla luciferase ofthis part come from psiCHECK-2 Vector.
+
Renilla Luciferase and Firefly Luciferase,as the control insert of this part both come from psiCHECK-2 Vector.
  
 
===References===
 
===References===
 +
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)

Revision as of 08:37, 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

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)