Difference between revisions of "Team:Czech Republic/Interlab study"

(→‎Used plasmids)
(→‎Materials and methods)
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5. Negative control (C-): K823008, in the pSB1C3 backbone
 
5. Negative control (C-): K823008, in the pSB1C3 backbone
 +
==Used strain==
 +
 +
E. coli E5alpha
 +
 +
==Used material==
 +
 +
1. LB-M agar plates with chloramphenicol
 +
 +
2. LB-M agar plates with ampicillin
 +
 +
3. 1,5 ml eppendorf tubes
 +
 +
4. 0,5 ml PCR tubes
 +
 +
5. 50 ml centrifuge conical base and rim tubes
 +
 +
6. NucleoSpin Plasmid DNA, RNA, and protein purification Kit
 +
 +
7. NucleoSpin Gel and PCR Clean-up Kit
 +
 +
8. LB-M medium with chloramphenicol
 +
 +
9. NaOH agarose gel and buffer
 +
 +
10. Sphero Rainbow Calibration Particles, 8 Peaks, 3.0-3.4
 +
 +
==Used methods (links)==
 +
 +
1. Transformation
 +
 +
2. Miniprep
 +
 +
3. Restriction digest
 +
 +
4. Ligation
 +
 +
5. NucleoSpin Gel Clean-up
 +
 +
6. NucleoSpin Plasmid DNA purification
 +
 +
7. Flow cytometry
 +
 +
==Used machines==
 +
 +
1. Incubator: Binder, ATP.line CB(E6), CO2 – Incubator with O2 control, model CB160, 230V
 +
 +
2. Orbita Shaker PSU-10i, 280 rpm, 360deg
 +
 +
3. Flow Cytometr C6
 +
 +
4. Thermo-Shaker with cooling for microtubes and PCR plates TS-100C
 +
 +
5. Centrifuge 5424/5424 R
 +
 +
==Used software==
 +
 +
1. CFlow Plus
 +
 +
2. Microsoft Excel
 +
 +
3. Sphero PMT QC Template
 +
 +
  
  

Revision as of 14:45, 3 September 2015

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Background

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Design

Materials and methods

Used plasmids

1. Device 1 (20K+21J): J23101 + I13504, in the pSB1C3 backbone

2. Device 2 (22A+21J): J23106 + I13504, in the pSB1C3 backbone

3. Device 3 (22K+21J): J23117 + I13504, in the pSB1C3 backbone

4. Positive control (C+): I20270, in the pSB1C3 backbone

5. Negative control (C-): K823008, in the pSB1C3 backbone

Used strain

E. coli E5alpha

Used material

1. LB-M agar plates with chloramphenicol

2. LB-M agar plates with ampicillin

3. 1,5 ml eppendorf tubes

4. 0,5 ml PCR tubes

5. 50 ml centrifuge conical base and rim tubes

6. NucleoSpin Plasmid DNA, RNA, and protein purification Kit

7. NucleoSpin Gel and PCR Clean-up Kit

8. LB-M medium with chloramphenicol

9. NaOH agarose gel and buffer

10. Sphero Rainbow Calibration Particles, 8 Peaks, 3.0-3.4

Used methods (links)

1. Transformation

2. Miniprep

3. Restriction digest

4. Ligation

5. NucleoSpin Gel Clean-up

6. NucleoSpin Plasmid DNA purification

7. Flow cytometry

Used machines

1. Incubator: Binder, ATP.line CB(E6), CO2 – Incubator with O2 control, model CB160, 230V

2. Orbita Shaker PSU-10i, 280 rpm, 360deg

3. Flow Cytometr C6

4. Thermo-Shaker with cooling for microtubes and PCR plates TS-100C

5. Centrifuge 5424/5424 R

Used software

1. CFlow Plus

2. Microsoft Excel

3. Sphero PMT QC Template




Protocols

Device Colony MEFL Mean of technical replicates SD Colony Mean of device SD Device
20K +21J (Device 1) Colony 1 I. 639549 657818 15504 664409 29185
II. 677453
III. 656453
Colony 2 I. 689717 702990 10067
II. 714084
III. 705169
Colony 3 I. 620331 632419 15053
II. 623286
III. 653639
22A+21J (Device 2) Colony 1 I. 481600 464304 12795 392603 50712
II. 460265
III. 451048
Colony 2 I. 346283 358070 9696
II. 357896
III. 370032
Colony 3 I. 354544 355433 4688
II. 350188
III. 361568
22K+21J (Device 3) Colony 1 I. 2635 2872 220 2229 454
II. 3165
III. 2814
Colony 2 I. 1786 1891 74
II. 1933
III. 1953
Colony 3 I. 1665 1926 204
II. 1948
III. 2163
Positive control Colony 1 I. 202361 196401 5853 196401 /
II. 188446
III. 198395
Negative control Colony 1 I. 224 213 8 213 /
II. 204
III. 211


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