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pipettetip_thermocycling_imaging

Thermocycling and imaging pipette tips

LabAutobox controls the temperature of 2 thermo blocks which the pipettes move into afterwards they can be imaged. To show the concept, the tips are incubating for only 5 seconds per block in the video.

The code works something like this:

  1. First you need to set the thermo block heaters to the temperature range that you want to run the thermocycling (ie., 94C denaturation, 60 extension).
//heatblockstemp block1 60 block2 100
M104 T0 S60
M104 T1 S100
  1. Then you just assemble the macro steps to form the routine, it would look something like this for one cycle
//thermocycler_imaging
//motion heatblock1 zh 0 row 1 F3000
G1Z0F3000
G1X34Y198.5Z0F3000_6
G1Z32F2000
G1Z42F400_30
//motion heatblock2 zh 0 row 1 F3000
G1Z0F3000
G1X36Y248Z0F3000_5
G1Z32F2000_2
G1Z42F400_180
G1Z0F3000
//motion PCRcam zh 0 row 1 F3000
G1Z0F3000
G1X5.5Y83.5F3000
G1Z46F3000
//camsnap 192.168.1.89_09302020070456_pipetteimaging   400 70
snap 192.168.1.89_400_70_09302020070456_pipetteimaging

This experiment tracks the volume per thermocycle (after 44 cycles). The aspiration conditions were as followed:

  1. Air gap 6µl
  2. Sample 6µl
  3. Air gap 4µl
  4. 94C 15 seconds
  5. 60C 30 seconds
cycle 1  cycle 2  cycle 3  cycle 4  cycle 5
cycle 6  cycle 7 {{ :347_50_thcycl1124104newpic.png?200 | cycle 8  cycle 9  cycle 10  cycle 11
cycle 12  cycle 13  cycle 14  cycle 15  cycle 16
cycle 17  cycle 18  cycle 19  cycle 20  cycle 21
cycle 22  cycle 23  cycle 24  cycle 25  cycle 26
cycle 27  cycle 28  cycle 29  cycle 30  cycle 31
cycle 32  cycle 33  cycle 34  cycle 35  cycle 36
cycle 37  cycle 38  cycle 39  cycle 40  cycle 41
cycle 42  cycle 43  cycle 44

Mouse beta-actin PCR

cyclesTemplate signalTemplate imageNo template signalNo template image
186.93589743589743 84.77777777777777
288.48148148148148 86.01754385964912
389.98148148148148 87.30952380952381
487.81111111111112 85.19047619047619
591.03030303030303 87.87719298245614
691.22619047619048 88.04545454545455
789.44827586206897 86.0
890.93827160493827 86.56666666666666
988.73076923076923 86.66666666666667
1092.92 92.11111111111111
1188.4367816091954 86.68253968253968
1287.72413793103448 98.33333333333333
1388.18666666666667 86.14035087719299
1490.09876543209876 87.06410256410257
1588.0 85.21794871794872
1688.46376811594203 85.42222222222222
1789.38333333333334 94.16666666666667
1893.52 90.11111111111111
1992.88888888888889 87.42666666666666
2091.51388888888889 87.07575757575758
2193.17391304347827 88.05797101449275
2293.81818181818181 88.73015873015873
2396.39393939393939 91.60606060606061
2494.98412698412699 86.88888888888889
2596.15 88.26984126984127
2696.2 88.41666666666667
2797.0701754385965 90.63333333333334
2898.14035087719299 90.54385964912281
2998.01754385964912 88.28070175438596
3097.79629629629629 91.43859649122807
31103.03703703703704 91.01960784313725
32100.3125 91.80392156862744
33103.04166666666667 92.3529411764706
34103.14583333333333 93.125
35105.8 93.4
36104.9375 95.53333333333333
37109.14285714285714 95.24444444444444
38104.85714285714286 97.97619047619048
39109.78571428571429 95.30952380952381
40108.51282051282051 96.5
41112.7948717948718 98.5
42116.57575757575758 102.02777777777777
43114.57575757575758 99.11111111111111
44116.87878787878788 102.21212121212122
45119.06060606060606 100.42424242424242
46119.93333333333334 101.15151515151516
47131.53333333333333 102.45454545454545
48123.23333333333333 106.33333333333333
49123.39393939393939 106.9
50124.66666666666667 111.26666666666667
pipettetip_thermocycling_imaging.txt · Last modified: 2021/08/16 15:22 (external edit)