User:Vinoo/2012 Shaker Test

Infors-Shaker-Platform.png MacConnell-Shaker-Platform.png
Growth Test Layout.png Cross Contamination Test Layout.png


Test 1 - August 1st, 2012

Prep

Part Used: BBa_J04450 in pSB1C3
Staging Plates: Single Colony Inoculations / Cross Contamination Test and Growth Test
Pin Tool: 5ul Pin Tool / Complete wash inbetween each inoculation
Plate Type: (8) Deep Well Plates (2ml)
Media: 1.8ml per well of LB Cm (Fresh LB and antibiotic)
Time: Incubation at 7:45pm - 11:45am

Shaker Location Test Seal Lid Temp Humidity RPM Comments
MacConnell Loc A CCT Micropore Yes 36C Unknown Unknown
MacConnell Loc D CCT Micropore Yes 36C Unknown Unknown
MacConnell Loc B Growth Micropore Yes 36C Unknown Unknown
MacConnell Loc C Growth Micropore Yes 36C Unknown Unknown
Infors 3mm Loc A2 CCT Micropore Yes 37C 70% 990rpm
Infors 3mm Loc D4 CCT Micropore Yes 37C 70% 990rpm
Infors 3mm Loc B5 Growth Micropore Yes 37C 70% 990rpm
Infors 3mm Loc C1 Growth Micropore Yes 37C 70% 990rpm
All plates in the Infors shaker had 3 out of 4 vinyl covers removed in order to accomodate plate lids.


Evaporation

Plate Before After
MacConnell Loc A 276.4g 274.46g
MacConnell Loc D 276.62g 274.63g
MacConnell Loc B 276.41g 274.54g
MacConnell Loc C 276.54g 274.51g
Infors 3mm Loc A2 276.66g 238.06g
Infors 3mm Loc D4 276.85g 245.62g
Infors 3mm Loc B5 277.31g 244.7g
Infors 3mm Loc C1 277.4g 225.54g
Evaporation results (weight after) for plates from Infors shaker must be disregarded due to culture spilling out during incubation.


Imaging

Growing Test

ShakerTest-8-1-12-Mac-Grow-LB.jpg ShakerTest-8-1-12-Mac-Grow-LC.jpg
ShakerTest-8-1-12-Infors-Grow-LA2.jpg ShakerTest-8-1-12-Infors-Grow-LD4.jpg

Top-Left: MacConnell Loc B
Top-Right: MacConnell Loc C
Bottom-Left: Infors 3mm Loc A2
Bottom-Right: Infors 3mm Loc D4


Cross Contamination Test

ShakerTest-8-1-12-Mac-CCT-LA.jpg ShakerTest-8-1-12-Mac-CCT-LD.jpg
ShakerTest-8-1-12-Infors-CCT-LB5.jpg ShakerTest-8-1-12-Infors-CCT-LC1.jpg

Top-Left: MacConnell Loc A
Top-Right: MacConnell Loc D
Bottom-Left: Infors 3mm Loc B5
Bottom-Right: Infors 3mm Loc C1


Conclusions

  • 990 rpm may be too high for 1.8ml of culture
  • removal of vinyl covers to accomodate lids allows for some freedom of movement for the base of the plate
  • micropore seal is not strong enough


Test 2 - August 3rd, 2012

Prep

Part Used: BBa_J04450 in pSB1C3
Staging Plate: Single Colony Inoculations / Cross Contamination Test
Pin Tool: 5ul Pin Tool / Complete wash inbetween each inoculation
Plate Type: (4) Deep Well Plates (2ml)
Media: 1.8ml per well of LB Cm (Fresh LB and antibiotic)
Time: Incubation at 5:30pm - 9:30am

Shaker Location Test Seal Lid Temp Humidity RPM Comments
MacConnell Loc A CCT Micropore Yes 36C Unknown Unknown
MacConnell Loc D CCT Micropore Yes 36C Unknown Unknown
Infors 3mm Loc A2 CCT Micropore No 37C 80% 700rpm
Infors 3mm Loc D4 CCT Micropore No 37C 80% 700rpm
Lids were not used on the plates in the Infors shaker so that all vinyl covers would be present to hold the base of the plate firmly


Evaporation

Plate Before After 1D 6D 12D
MacConnell Loc A 276.03g 274.67g 1.71 ml 1.69 ml 1.68 ml
MacConnell Loc D 276.07g 274.89g 1.7 ml 1.695 ml 1.71 ml
Infors 3mm Loc A2 275.96g 270.83g 1.67 ml 1.64 ml 1.625 ml
Infors 3mm Loc D4 276.1g 271.43g 1.65 ml 1.68 ml 1.62 ml


Cell Culture OD

(600nm Abs Values)

Plate 1C 3A 6D 9G 12D 12H
MacConnell Loc A 2.22 2.5 2.5 1.92 2.52 2.05
MacConnell Loc D 1.66 1.45 1.45 1.85 1.58 1.83
Infors 3mm Loc A2 2.31 2.47 2.27 2.34 2.64 2.39
Infors 3mm Loc D4 2.4 1.28 2.31 2.13 2.72 2.42

Imaging

ShakerTest-8-3-12-Mac-CCT-LA.jpg ShakerTest-8-3-12-Mac-CCT-LD.jpg
ShakerTest-8-3-12-Infors-CCT-LA2.jpg ShakerTest-8-3-12-Infors-CCT-LD4.jpg

Top-Left: MacConnell Loc A
Top-Right: MacConnell Loc D
Bottom-Left: Infors 3mm Loc A2
Bottom-Right: Infors 3mm Loc D4

Contamination in wells 4C, 7D, 8C in the Infors 3mm Loc D4 plate, and in 10C in the MacConnell Loc A plate.


Conclusions

  • 700 rpm grows culture just as well as the MacConnell, but more consistently
  • contamination in 3 of the wells
    • aerosol bacteria enters a well, through the micropore, or if the seal on the micropore breaks
    • liquid culture enters a well, through the micropore, or if the seal on the micropore breaks
    • pin tool was contaminated


Test 3 - August 13th, 2012

  • Plates were prepared with LB Broth (Cm) and blue food dye
  • No plate lids used
  • All vinyl covers used to firmly hold plate base in place
  • Plates were spun at the specified RPM for 1 hour
  • Micropore was not changed between tests
  • Plates were weighed with their micropore on
  • Plates were imaged with micropore on
  • After last test 990rpm micropore was removed and checked for any blue media spots/splashes
Volume Location Weight 700rpm Weight After 800rpm Weight After 900rpm Weight After 990rpm Weight After
1.8ml Infors 3mm - C1 280.35g No 280.38g No 279.98g No 279.37g No 278.69g
1.7ml Infors 3mm - B5 270.18g No 270.33g No 270.11g No 269.70g No 269.25
1.6ml Infors 3mm - D4 260.46g No 260.58g No 260.28g No 259.87g No 259.39g
1.5ml Infors 3mm - B2 250.73g No 250.78g No 250.48g No 250.07g No 249.62g

Test 4 - August 14th, 2012

  • Plates were prepared with LB Broth (Cm) and blue food dye
  • Plate lids were used
  • 3 out of 4 covers removed
  • Plates were spun at the specified RPM for 1 hour
  • Micropore was not changed between tests
  • Plates were weighed with their micropore on
  • Plates were imaged with micropore on
  • After last test 990rpm micropore was removed and checked for any blue media spots/splashes
Volume Location Weight 700rpm Weight After 800rpm Weight After 900rpm Weight After 990rpm Weight After
1.8ml Infors 3mm - C1 280.45g No 280.44g No 280.34g No 279.98g Yes 278.45g
1.7ml Infors 3mm - B5 270.84g No 270.86g No 270.77g Yes* 270.22g Yes 267.64
1.6ml Infors 3mm - D4 261.15g No 261.18g No 261.1g No 260.86g Yes* 260.45g
1.5ml Infors 3mm - B2 250.81g No 250.83g No 250.68g No 250.3g No 248.89g