Monday, November 10, 2014

Dictyostelium Predation Assay for transposon mutagensis of pDR2 Part(1)

Date: Nov./1/2014
The protocol in the link
Plate name :  pDR 2/Tetra. transposon mutagensis.
 The result: after 4 days incubation at room temp. in a dark place
Plaques in Kleb. plate=35 plaques in dilution 1/1000 (-3)

Plaque forming unit pfu =# plaque X dilution factor/volume of inoculum
                                       35 X10^3/10 = 3.5 X10^3
μl of  8 Dicty. dilutions that contain(7.0 X 10^3, 7.0 X 10^2, 7.0 X 10 ,7.0 , 0.7,....) Dicty. cells were used on sm2 agar in four replicate for each dilution.
The result after 4 days :
Controls: 

  • pDR2plague :( 0, -1, -2 ) 
  •  cosmid control.plague(0,-1,-2,-3,-4)
  • Clones shows sensitivity to Dictyostelium predation:
plague (0,-1,-2,-3)
A7     named: pDR2  tn 1
A8     named: pDR2  tn 2
A12   named: pDR2  tn 3
B3     named: pDR2  tn 41
C4     named: pDR2  tn 5 
D3     named: pDR2  tn 6
D11   named: pDR2  tn 7 
E2     named: pDR2   tn 8 
E5     named: pDR2   tn 9
E6     named: pDR2   tn 10 
E8     named: pDR 2  tn 11


plague (0,-1,-2,-3,-4) 
E4 named: pDR2tn12

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