====== 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. 

{{youtube>yU6GE9LnJxI?&rel=0 | Pipette tip thermocycling example}}

The code works something like this:

  - 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). 
<code>
//heatblockstemp block1 60 block2 100
M104 T0 S60
M104 T1 S100
</code>
  - Then you just assemble the macro steps to form the routine, it would look something like this for one cycle
<code>
//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

</code>


This experiment tracks the volume per thermocycle (after 44 cycles). The aspiration conditions were as followed:
  - Air gap 6µl
  - Sample 6µl
  - Air gap 4µl
  - 94C 15 seconds
  - 60C 30 seconds


| cycle 1 {{ :347_50_thcycl5124824newpic.png?200 | }} | cycle 2 {{ :347_50_thcycl6124936newpic.png?200 | }} | cycle 3 {{ :347_50_thcycl7125049newpic.png?200 | }} | cycle 4 {{ :347_50_thcycl8125202newpic.png?200 | }} | cycle 5 {{ :347_50_thcycl9125315newpic.png?200 | }} |
| cycle 6 {{ :347_50_thcycl10125427newpic.png?200 | cycle 7 {{ :347_50_thcycl1124104newpic.png?200 | }} | cycle 8 {{ :347_50_thcycl12125653newpic.png?200 | }} | cycle 9 {{ :347_50_thcycl13125805newpic.png?200 | }} | cycle 10 {{ :347_50_thcycl14125918newpic.png?200 | }} | cycle 11 {{ :347_50_thcycl15130031newpic.png?200 | }} |
| cycle 12 {{ :347_50_thcycl16130143newpic.png?200 | }} | cycle 13 {{ :347_50_thcycl17130256newpic.png?200 | }} | cycle 14 {{ :347_50_thcycl18130409newpic.png?200 | }} | cycle 15 {{ :347_50_thcycl19130522newpic.png?200 | }} | cycle 16 {{ :347_50_thcycl20130634newpic.png?200 | }} |
| cycle 17 {{ :347_50_thcycl21130747newpic.png?200 | }} | cycle 18 {{ :347_50_thcycl2124229newpic.png?200 | }} | cycle 19 {{ :347_50_thcycl22130900newpic.png?200 | }} | cycle 20 {{ :347_50_thcycl23131012newpic.png?200 | }} | cycle 21 {{ :347_50_thcycl24131125newpic.png?200 | }} |
| cycle 22 {{ :347_50_thcycl25131238newpic.png?200 | }} | cycle 23 {{ :347_50_thcycl26131350newpic.png?200 | }} | cycle 24 {{ :347_50_thcycl27131503newpic.png?200 | }} | cycle 25 {{ :347_50_thcycl28131616newpic.png?200 | }} | cycle 26 {{ :347_50_thcycl29131728newpic.png?200 | }} |
| cycle 27 {{ :347_50_thcycl30131841newpic.png?200 | }} | cycle 28 {{ :347_50_thcycl31131954newpic.png?200 | }} | cycle 29 {{ :347_50_thcycl3124354newpic.png?200 | }} | cycle 30 {{ :347_50_thcycl32132107newpic.png?200 | }} | cycle 31 {{ :347_50_thcycl33132219newpic.png?200 | }} |
| cycle 32 {{ :347_50_thcycl34132332newpic.png?200 | }} | cycle 33 {{ :347_50_thcycl35132445newpic.png?200 | }} | cycle 34 {{ :347_50_thcycl36132557newpic.png?200 | }} | cycle 35 {{ :347_50_thcycl37132710newpic.png?200 | }} | cycle 36 {{ :347_50_thcycl38132823newpic.png?200 | }} |
| cycle 37 {{ :347_50_thcycl39132936newpic.png?200 | }} | cycle 38 {{ :347_50_thcycl40133048newpic.png?200 | }} | cycle 39 {{ :347_50_thcycl41133201newpic.png?200 | }} | cycle 40 {{ :347_50_thcycl4124519newpic.png?200 | }} | cycle 41 {{ :347_50_thcycl42133314newpic.png?200 | }} |
| cycle 42 {{ :347_50_thcycl43133426newpic.png?200 | }} | cycle 43 {{ :347_50_thcycl44133539newpic.png?200 | }} | cycle 44 {{ :347_50_thcycl45133652newpic.png?200 | }} |


----

Mouse beta-actin PCR


{{ :bdev:firstpipettepcrgraph.png?600 |}}


|cycles|Template signal|Template image|No template signal|No template image|
|1|86.93589743589743| {{ :bdev:template_350_60_thermocycle0pos56.5133827.png |}} |84.77777777777777| {{ :bdev:notemplate_350_60_thermocycle0pos54.5133806.png |}} |
|2|88.48148148148148| {{ :bdev:template_350_60_thermocycle1pos58134036.png |}} |86.01754385964912| {{ :bdev:notemplate_350_60_thermocycle1pos54.5133959.png |}} |
|3|89.98148148148148| {{ :bdev:template_350_60_thermocycle2pos58134229.png |}} |87.30952380952381| {{ :bdev:notemplate_350_60_thermocycle2pos58134229.png |}} |
|4|87.81111111111112| {{ :bdev:template_350_60_thermocycle3pos56.5134407.png |}} |85.19047619047619| {{ :bdev:notemplate_350_60_thermocycle3pos56.5134407.png |}} |
|5|91.03030303030303| {{ :bdev:template_350_60_thermocycle4pos57.5134610.png |}} |87.87719298245614| {{ :bdev:notemplate_350_60_thermocycle4pos57.5134610.png |}} |
|6|91.22619047619048| {{ :bdev:template_350_60_thermocycle5pos56134748.png |}} |88.04545454545455| {{ :bdev:notemplate_350_60_thermocycle5pos57134759.png |}} |
|7|89.44827586206897| {{ :bdev:template_350_60_thermocycle6pos56134941.png |}} |86.0| {{ :bdev:notemplate_350_60_thermocycle6pos53134910.png |}} |
|8|90.93827160493827| {{ :bdev:template_350_60_thermocycle7pos56.5135140.png |}} |86.56666666666666| {{ :bdev:notemplate_350_60_thermocycle7pos54.5135119.png |}} |
|9|88.73076923076923| {{ :bdev:template_350_60_thermocycle8pos56.5135333.png |}} |86.66666666666667| {{ :bdev:notemplate_350_60_thermocycle8pos58135349.png |}} |
|10|92.92| {{ :bdev:template_350_60_thermocycle9pos57135532.png |}} |92.11111111111111| {{ :bdev:notemplate_350_60_thermocycle9pos54.5135506.png |}} |
|11|88.4367816091954| {{ :bdev:template_350_60_thermocycle10pos54.5135705.png |}} |86.68253968253968| {{ :bdev:notemplate_350_60_thermocycle10pos56135721.png |}} |
|12|87.72413793103448| {{ :bdev:template_350_60_thermocycle11pos55135904.png |}} |98.33333333333333| {{ :bdev:notemplate_350_60_thermocycle11pos58135935.png |}} |
|13|88.18666666666667| {{ :bdev:template_350_60_thermocycle12pos56140108.png |}} |86.14035087719299| {{ :bdev:notemplate_350_60_thermocycle12pos56140108.png |}} |
|14|90.09876543209876| {{ :bdev:template_350_60_thermocycle13pos55.5140256.png |}} |87.06410256410257| {{ :bdev:notemplate_350_60_thermocycle13pos55.5140256.png |}} |
|15|88.0| {{ :bdev:template_350_60_thermocycle14pos55140444.png |}} |85.21794871794872| {{ :bdev:notemplate_350_60_thermocycle14pos55140444.png |}} |
|16|88.46376811594203| {{ :bdev:template_350_60_thermocycle15pos56.5140653.png |}} |85.42222222222222| {{ :bdev:notemplate_350_60_thermocycle15pos56.5140653.png |}} |
|17|89.38333333333334| {{ :bdev:template_350_60_thermocycle16pos57140851.png |}} |94.16666666666667| {{ :bdev:notemplate_350_60_thermocycle16pos58140902.png |}} |
|18|93.52| {{ :bdev:template_350_60_thermocycle17pos55141024.png |}} |90.11111111111111| {{ :bdev:notemplate_350_60_thermocycle17pos55141024.png |}} |
|19|92.88888888888889| {{ :bdev:template_350_60_thermocycle18pos55.5141222.png |}} |87.42666666666666| {{ :bdev:notemplate_350_60_thermocycle18pos55.5141222.png |}} |
|20|91.51388888888889| {{ :bdev:template_350_60_thermocycle19pos55.5141416.png |}} |87.07575757575758| {{ :bdev:notemplate_350_60_thermocycle19pos55141410.png |}} |
|21|93.17391304347827| {{ :bdev:template_350_60_thermocycle20pos55.5141615.png |}} |88.05797101449275| {{ :bdev:notemplate_350_60_thermocycle20pos55.5141615.png |}} |
|22|93.81818181818181| {{ :bdev:template_350_60_thermocycle21pos56141814.png |}} |88.73015873015873| {{ :bdev:notemplate_350_60_thermocycle21pos56141814.png |}} |
|23|96.39393939393939| {{ :bdev:template_350_60_thermocycle22pos55.5142002.png |}} |91.60606060606061| {{ :bdev:notemplate_350_60_thermocycle22pos55141957.png |}} |
|24|94.98412698412699| {{ :bdev:template_350_60_thermocycle23pos56.5142206.png |}} |86.88888888888889| {{ :bdev:notemplate_350_60_thermocycle23pos56.5142206.png |}} |
|25|96.15| {{ :bdev:template_350_60_thermocycle24pos56.5142359.png |}} |88.26984126984127| {{ :bdev:notemplate_350_60_thermocycle24pos55.5142348.png |}} |
|26|96.2| {{ :bdev:template_350_60_thermocycle25pos56142547.png |}} |88.41666666666667| {{ :bdev:notemplate_350_60_thermocycle25pos56142547.png |}} |
|27|97.0701754385965| {{ :bdev:template_350_60_thermocycle26pos56.5142745.png |}} |90.63333333333334| {{ :bdev:notemplate_350_60_thermocycle26pos54.5142725.png |}} |
|28|98.14035087719299| {{ :bdev:template_350_60_thermocycle27pos56142934.png |}} |90.54385964912281| {{ :bdev:notemplate_350_60_thermocycle27pos56142934.png |}} |
|29|98.01754385964912| {{ :bdev:template_350_60_thermocycle28pos57143137.png |}} |88.28070175438596| {{ :bdev:notemplate_350_60_thermocycle28pos55.5143122.png |}} |
|30|97.79629629629629| {{ :bdev:template_350_60_thermocycle29pos56143320.png |}} |91.43859649122807| {{ :bdev:notemplate_350_60_thermocycle29pos55.5143315.png |}} |
|31|103.03703703703704| {{ :bdev:template_350_60_thermocycle30pos57.5143859.png |}} |91.01960784313725| {{ :bdev:notemplate_350_60_thermocycle30pos56143844.png |}} |
|32|100.3125| {{ :bdev:template_350_60_thermocycle31pos56.5144042.png |}} |91.80392156862744| {{ :bdev:notemplate_350_60_thermocycle31pos56144037.png |}} |
|33|103.04166666666667| {{ :bdev:template_350_60_thermocycle32pos57144241.png |}} |92.3529411764706| {{ :bdev:notemplate_350_60_thermocycle32pos56144230.png |}} |
|34|103.14583333333333| {{ :bdev:template_350_60_thermocycle33pos57144434.png |}} |93.125| {{ :bdev:notemplate_350_60_thermocycle33pos55.5144418.png |}} |
|35|105.8| {{ :bdev:template_350_60_thermocycle34pos57.5144633.png |}} |93.4| {{ :bdev:notemplate_350_60_thermocycle34pos55.5144612.png |}} |
|36|104.9375| {{ :bdev:template_350_60_thermocycle35pos57.5144826.png |}} |95.53333333333333| {{ :bdev:notemplate_350_60_thermocycle35pos56144810.png |}} |
|37|109.14285714285714| {{ :bdev:template_350_60_thermocycle36pos57145014.png |}} |95.24444444444444| {{ :bdev:notemplate_350_60_thermocycle36pos55144953.png |}} |
|38|104.85714285714286| {{ :bdev:template_350_60_thermocycle37pos56145157.png |}} |97.97619047619048| {{ :bdev:notemplate_350_60_thermocycle37pos56145157.png |}} |
|39|109.78571428571429| {{ :bdev:template_350_60_thermocycle38pos57.5145406.png |}} |95.30952380952381| {{ :bdev:notemplate_350_60_thermocycle38pos55.5145345.png |}} |
|40|108.51282051282051| {{ :bdev:template_350_60_thermocycle39pos57.5145559.png |}} |96.5| {{ :bdev:notemplate_350_60_thermocycle39pos55.5145538.png |}} |
|41|112.7948717948718| {{ :bdev:template_350_60_thermocycle40pos57145747.png |}} |98.5| {{ :bdev:notemplate_350_60_thermocycle40pos56150118.png |}} |
|42|116.57575757575758| {{ :bdev:template_350_60_thermocycle41pos57.5150327.png |}} |102.02777777777777| {{ :bdev:notemplate_350_60_thermocycle41pos56150312.png |}} |
|43|114.57575757575758| {{ :bdev:template_350_60_thermocycle42pos57150516.png |}} |99.11111111111111| {{ :bdev:notemplate_350_60_thermocycle42pos56.5150510.png |}} |
|44|116.87878787878788| {{ :bdev:template_350_60_thermocycle43pos57.5150714.png |}} |102.21212121212122| {{ :bdev:notemplate_350_60_thermocycle43pos55.5150653.png |}} |
|45|119.06060606060606| {{ :bdev:template_350_60_thermocycle44pos57.5150907.png |}} |100.42424242424242| {{ :bdev:notemplate_350_60_thermocycle44pos55150841.png |}} |
|46|119.93333333333334| {{ :bdev:template_350_60_thermocycle45pos57.5151101.png |}} |101.15151515151516| {{ :bdev:notemplate_350_60_thermocycle45pos55.5151040.png |}} |
|47|131.53333333333333| {{ :bdev:template_350_60_thermocycle46pos57.5151254.png |}} |102.45454545454545| {{ :bdev:notemplate_350_60_thermocycle46pos57151249.png |}} |
|48|123.23333333333333| {{ :bdev:template_350_60_thermocycle47pos58151452.png |}} |106.33333333333333| {{ :bdev:notemplate_350_60_thermocycle47pos56151432.png |}} |
|49|123.39393939393939| {{ :bdev:template_350_60_thermocycle48pos58151646.png |}} |106.9| {{ :bdev:notemplate_350_60_thermocycle48pos56151625.png |}} |
|50|124.66666666666667| {{ :bdev:template_350_60_thermocycle49pos57.5151834.png |}} |111.26666666666667| {{ :bdev:notemplate_350_60_thermocycle49pos55.5151813.png |}} |