Dictyostelium Predation Assay for transposon mutagensis of pDR2 Part(2)

Date: 10/1/2014
The protocol in the link
Plate name :  pDR 2/Kana. P1 & 4 , transposon mutagensis.
 The result: after 4 days incubation at room temp. in a dark place
 Plaques in Kleb. plate=1 plaques in dilution 1/1000 (-3)

Plaque forming unit pfu =# plaque X dilution factor/volume of inoculum
                                      =1.0 X10^3

2 μl of  8 Dicty. dilutions that contain(2.0 X 10^2, 2.0 X 10^1, 2 , 0.2,....)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)
Clones shows sensitivity to Dictyostelium predation:
plague (0,-1,-2,-3)

Dictyostelium Predation Assay for transposon mutagensis of pDR1 Part(4)

Date: Nov./10/2014
The protocol in the link
Plate name :collection of 3 paltes(contain 10 +20+10 colonies), pDR1/kana. transposon mutagensis.
 The result: after 4 days incubation at room temp. in a dark place

Plaques in Kleb. plate=1 plaques in dilution 1/1000 (-3)

Plaque forming unit pfu =# plaque X dilution factor/volume of inoculum
                                      =1.0 X10^3
2 μl of  8 Dicty. dilutions that contain(2.0 X 10^2, 2.0 X 10^1, 2 , 0.2,....) Dicty. cells were used on sm2 agar in four replicate for each dilution.
The result after 4 days :
Controls: 

• pDR1plague :(0) 
•  cosmid control.plague(0,-1,-2)
• Clones shows sensitivity to Dictyostelium predation:

plague (0,-1,-2) 
9-2A8 named: pDR1tn9

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
2 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