User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/04/22: Difference between revisions

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*Ligation of construct KAH182-KAH201 and mammalian vector V0200 and cloning in DH5α-T..
*Ligation of construct KAH182-KAH201 and mammalian vector V0200 and cloning in DH5α-T..
KAH182 = BBa_S04739 and KAH201 = BBa_S04745 and V0200 = BBa_J176121.   
KAH182 = BBa_S04739 and KAH201 = BBa_S04745 and V0200 = BBa_J176121.   
*Assembly strategy: The backbone KAH201(S/P)/3403 , the insert is KAH182(X/P)/1675.
*Assembly strategy: The backbone V0200(N/S)/4442 , the insert is KAH182-KAH201(N/S)/1901.




Revision as of 15:44, 22 April 2013

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Date

Building a Reporter Gene to Determine Chromatin Protein Function (Ligation into the mammalian vector)


  • Ligation of construct KAH182-KAH201 and mammalian vector V0200 and cloning in DH5α-T..

KAH182 = BBa_S04739 and KAH201 = BBa_S04745 and V0200 = BBa_J176121.

  • Assembly strategy: The backbone V0200(N/S)/4442 , the insert is KAH182-KAH201(N/S)/1901.


Digests


(1,2) KAH182, 1675 bp, Cut with Xba1 and Pst1, (3,4) KAH201, 3403 bp, Cut with Spe1 and Pst1
Plasmid DNA 25.00 μl
NotI 1.0 μl
SpeI 1.0 μl
10x FastDigest buffer + green loading dye 3.00 μl
dH2O 0.00 μl
  30.0 μl total
Incubate at 37°C for 10 minutes.
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