Thursday, October 30, 2014

Dictyostelium Predation Assay for transposon mutagensis of pDR1 Part(2)

Date: Oct./25/2014
The protocol in the link
Plate 4, pDR1/kana. transposon mutagensis.
 The result: after 4 days incubation at room temp. in a dark place
Plaques in Kleb. plate=23 plaques in dilution 1/1000 (-3)

Plaque forming unit pfu =# plaque X dilution factor/volume of inoculum
                                       23 X10^3/10 = 2.3 X10^3
μl of  8 Dicty. dilutions that contain(4.6 X 10^3, 4.6 X 10^2,4.6 X 10 ,4.6 , 0.46 ,....) Dicty. cells were used on sm2 agar in four replicate for each dilution.
The result after 4 days :
Controls: 

  • pDR1plague :(0,-1,-2) 
  •  cosmid control.plague(0,-1,-2,-3,-4)
  • Clones shows sensitivity to Dictyostelium predation:
plague (0,-1,-2,-3) 
6-A7 named: pDR1tn6 - small palques-

plague(0,-1,-2,-3,-4)
7-A6   named:   pDR1tn6
The result after 5 days incubation look like same 

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