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

From OpenWetWare
Jump to: navigation, search
(Date)
(Date)
Line 12: Line 12:
  
  
*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 V0200(N/S)/4442 , the insert is KAH182-KAH201(N/S)/1901.
 
*Assembly strategy: The backbone V0200(N/S)/4442 , the insert is KAH182-KAH201(N/S)/1901.
  
Line 20: Line 20:
  
  
[[Image:KAH182-KAH201-V0200-S-N Digestion.jpg|350px| (1) KAH182-KAH201, 1901 bp, (2) V0200, 4442 bp, Both 1 and 2 are cut with Spe1 and Not11]]
+
[[Image:KAH182-KAH201-V0200-S-N Digestion.jpg|thumb|350px| (1) KAH182-KAH201, 1901 bp, (2) V0200, 4442 bp, Both 1 and 2 are cut with Spe1 and Not11]]
  
 
{| {{table}} cellspacing="3"  width=350px  
 
{| {{table}} cellspacing="3"  width=350px  

Revision as of 15:58, 22 April 2013

Owwnotebook icon.pngPcTF Subcloning in E. coli <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

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) KAH182-KAH201, 1901 bp, (2) V0200, 4442 bp, Both 1 and 2 are cut with Spe1 and Not11
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.