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 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

cycles	Template signal	Template image	No template signal	No template image
1	86.93589743589743		84.77777777777777
2	88.48148148148148		86.01754385964912
3	89.98148148148148		87.30952380952381
4	87.81111111111112		85.19047619047619
5	91.03030303030303		87.87719298245614
6	91.22619047619048		88.04545454545455
7	89.44827586206897		86.0
8	90.93827160493827		86.56666666666666
9	88.73076923076923		86.66666666666667
10	92.92		92.11111111111111
11	88.4367816091954		86.68253968253968
12	87.72413793103448		98.33333333333333
13	88.18666666666667		86.14035087719299
14	90.09876543209876		87.06410256410257
15	88.0		85.21794871794872
16	88.46376811594203		85.42222222222222
17	89.38333333333334		94.16666666666667
18	93.52		90.11111111111111
19	92.88888888888889		87.42666666666666
20	91.51388888888889		87.07575757575758
21	93.17391304347827		88.05797101449275
22	93.81818181818181		88.73015873015873
23	96.39393939393939		91.60606060606061
24	94.98412698412699		86.88888888888889
25	96.15		88.26984126984127
26	96.2		88.41666666666667
27	97.0701754385965		90.63333333333334
28	98.14035087719299		90.54385964912281
29	98.01754385964912		88.28070175438596
30	97.79629629629629		91.43859649122807
31	103.03703703703704		91.01960784313725
32	100.3125		91.80392156862744
33	103.04166666666667		92.3529411764706
34	103.14583333333333		93.125
35	105.8		93.4
36	104.9375		95.53333333333333
37	109.14285714285714		95.24444444444444
38	104.85714285714286		97.97619047619048
39	109.78571428571429		95.30952380952381
40	108.51282051282051		96.5
41	112.7948717948718		98.5
42	116.57575757575758		102.02777777777777
43	114.57575757575758		99.11111111111111
44	116.87878787878788		102.21212121212122
45	119.06060606060606		100.42424242424242
46	119.93333333333334		101.15151515151516
47	131.53333333333333		102.45454545454545
48	123.23333333333333		106.33333333333333
49	123.39393939393939		106.9
50	124.66666666666667		111.26666666666667