Selected Cell
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plate
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| RT CPB stages 96 well plate | ||||||||||||||||||||||||||||
| IDs | ||||||||||||||||||||||||||||
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | RT MIX | 33 | overhead: | 0.15 | |||||||||||||
| A | RI009 | RI015 | RI025 | RI035 | RI041 | 1 reaction | reactions: | |||||||||||||||||||||
| B | RI010 | RI016 | RI027 | RI037 | RT buffer | 2.5 | 94.87499999999999 | |||||||||||||||||||||
| C | RI234 | RI018 | RI028 | RI241 | dNTP_mix | 1 | 37.949999999999996 | |||||||||||||||||||||
| D | RI237 | RI019 | RI030 | RI242 | random primers | 2.5 | 94.87499999999999 | |||||||||||||||||||||
| E | RI011 | RI020 | RI031 | RI243 | water | 4.25 | 161.2875 | |||||||||||||||||||||
| F | RI012 | RI021 | RI032 | RI038 | Rnase inhibitor | 1 | 37.949999999999996 | |||||||||||||||||||||
| G | RI013 | RI023 | RI033 | RI039 | RT enzyme | 1.25 | 47.43749999999999 | |||||||||||||||||||||
| H | RI014 | RI024 | RI034 | RI040 | 12.5 | 474.37499999999994 | ||||||||||||||||||||||
| qPCR CPB stages | ||||||||||||||||||||||||||||
| sample names | ||||||||||||||||||||||||||||
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |||||
| A | RI009 | RI015 | RI025 | RI035 | RI041 | A | egg early 3 | neonatal 2 | 3rd larva 3 | 4th late larva 4 | adult 4 | |||||||||||||||||
| B | RI010 | RI016 | RI027 | RI037 | RI001 | B | egg early 4 | neonatal 3 | 4th early larva 1 | pupa 1 | foregut | |||||||||||||||||
| C | RI234 | RI018 | RI028 | RI241 | RI002 | C | eggs early 2 | 2nd larva 2 | 4th early larva 2 | pupa 2 | midgut | |||||||||||||||||
| D | RI237 | RI019 | RI030 | RI242 | RI003 | D | eggs early 3 | 2nd larva 3 | 4th early larva 3 | pupa 3 | hundgut | |||||||||||||||||
| E | RI011 | RI020 | RI031 | RI243 | RI004 | E | egg late 1 | 2nd larva 4 | 4th early larva 4 | pupa 4 | haemolymph | |||||||||||||||||
| F | RI012 | RI021 | RI032 | RI038 | RI005 | F | egg late 2 | 2nd larva 5 | 4th late larva 1 | adult 1 | carcass | |||||||||||||||||
| G | RI013 | RI023 | RI033 | RI039 | RI006 | G | egg late 3 | 3rd larva 1 | 4th late larva 2 | adult 2 | legs | |||||||||||||||||
| H | RI014 | RI024 | RI034 | RI040 | H | neonatal 1 | 3rd larva 2 | 4th late larva 3 | adult 3 | |||||||||||||||||||
| REF | ||||||||||||||||||||||||||||
| qPCR plates and genes: | taqMan | 18S | alas | mesh | JHBP_TqMn | |||||||||||||||||||||||
| SYBR | LdRP4 | JHBP-like | ||||||||||||||||||||||||||
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| SampleID | SampleName | SampleName_Generated | ng/ul totRNA | ul totRNA | ug totRNA | RT Vol | H2O Vol | RT ng | |
| RI001 | foregut | larva_4th_instar_foregut-1 | NA (cDNA!) | NA (cDNA!) | NA (cDNA!) | NA | NA | NA | |
| RI002 | midgut | larva_4th_instar_midgut-1 | NA (cDNA!) | NA (cDNA!) | NA (cDNA!) | NA | NA | NA | |
| RI003 | hindgut | larva_4th_instar_hindgut-1 | NA (cDNA!) | NA (cDNA!) | NA (cDNA!) | NA | NA | NA | |
| RI004 | haemolymph | larva_4th_instar_haemolymph-1 | NA (cDNA!) | NA (cDNA!) | NA (cDNA!) | NA | NA | NA | |
| RI005 | carcass | larva_4th_instar_carcass-1 | NA (cDNA!) | NA (cDNA!) | NA (cDNA!) | NA | NA | NA | |
| RI006 | legs | larva_4th_instar_legs-1 | NA (cDNA!) | NA (cDNA!) | NA (cDNA!) | NA | NA | NA | |
| RI009 | egg_early-3 | egg_early_whole_egg-3 | 13 | 12 | 0.156 | 12.5 | 0 | 162.5 | |
| RI010 | egg_early-4 | egg_early_whole_egg-4 | 14 | 12 | 0.168 | 12.5 | 0 | 175 | |
| RI011 | egg_late-1 | egg_late_whole_egg-1 | 184 | 12 | 2.208 | 8 | 4.5 | 1472 | |
| RI012 | egg_late-2 | egg_late_whole_egg-2 | 114 | 12 | 1.368 | 12.5 | 0 | 1425 | |
| RI013 | egg_late-3 | egg_late_whole_egg-3 | 143 | 12 | 1.716 | 8 | 4.5 | 1144 | |
| RI014 | larva_neonatal-1 | larva_neonatal_whole_larva-1 | 58 | 12 | 0.696 | 12.5 | 0 | 725 | |
| RI015 | larva_neonatal-2 | larva_neonatal_whole_larva-2 | 70 | 12 | 0.84 | 12.5 | 0 | 875 | |
| RI016 | larva_neonatal-3 | larva_neonatal_whole_larva-3 | 65 | 12 | 0.78 | 12.5 | 0 | 812.5 | |
| RI018 | larva_2nd-2 | larva_2th_instar_gut-2 | 147 | 12 | 1.764 | 8 | 4.5 | 1176 | |
| RI019 | larva_2nd-3 | larva_2th_instar_gut-3 | 312 | 12 | 3.744 | 5 | 7.5 | 1560 | |
| RI020 | larva_2nd-4 | larva_2th_instar_gut-4 | 126 | 12 | 1.512 | 8 | 4.5 | 1008 | |
| RI021 | larva_2nd-5 | larva_2th_instar_gut-5 | 214 | 12 | 2.568 | 5 | 7.5 | 1070 | |
| RI023 | larva_3rd-1 | larva_3rd_instar_gut-1 | 36 | 22 | 0.792 | 12.5 | 0 | 450 | |
| RI024 | larva_3rd-2 | larva_3rd_instar_gut-2 | 169 | 22 | 3.718 | 8 | 4.5 | 1352 | |
| RI025 | larva_3rd-3 | larva_3rd_instar_gut-3 | 94 | 22 | 2.068 | 12.5 | 0 | 1175 | |
| RI027 | larva_4th_early-1 | larva_4th_instar_early_midgut-1 | 785 | 22 | 17.27 | 2 | 10.5 | 1570 | |
| RI028 | larva_4th_early-2 | larva_4th_instar_early_midgut-2 | 153 | 22 | 3.366 | 8 | 4.5 | 1224 | |
| RI030 | larva_4th_early-4 | larva_4th_instar_early_midgut-4 | 118 | 22 | 2.596 | 12.5 | 0 | 1475 | |
| RI031 | larva_4th_early-5 | larva_4th_instar_early_midgut-5 | 327 | 22 | 7.194 | 5 | 7.5 | 1635 | |
| RI032 | larva_4th_late-1 | larva_4th_instar_late_midgut-1 | 178 | 22 | 3.916 | 8 | 4.5 | 1424 | |
| RI033 | larva_4th_late-2 | larva_4th_instar_late_midgut-2 | 196 | 22 | 4.312 | 8 | 4.5 | 1568 | |
| RI034 | larva_4th_late-3 | larva_4th_instar_late_midgut-3 | 212 | 22 | 4.664 | 8 | 4.5 | 1696 | |
| RI035 | larva_4th_late-4 | larva_4th_instar_late_midgut-4 | 280 | 22 | 6.16 | 5 | 7.5 | 1400 | |
| RI037 | pupa-1 | pupa_gut-1 | 281 | 12 | 3.372 | 5 | 7.5 | 1405 | |
| RI038 | adult_1 | adult_midgut-1 | 152 | 22 | 3.344 | 8 | 4.5 | 1216 | |
| RI039 | adult_2 | adult_midgut-2 | 276 | 22 | 6.072 | 5 | 7.5 | 1380 | |
| RI040 | adult_3 | adult_midgut-3 | 839 | 22 | 18.458 | 2 | 10.5 | 1678 | |
| RI041 | adult_4 | adult_midgut-4 | 369 | 22 | 8.118 | 5 | 7.5 | 1845 | |
| RI0234 | eggs_early_1 | egg_early_whole_egg-1 | 276 | 14 | 3.864 | 5 | 7.5 | 1380 | |
| RI0237 | eggs_early_4 | egg_early_whole_egg-4 | 42 | 14 | 0.588 | 12.5 | 0 | 525 | |
| RI0241 | pupa_gut_2 | pupa_gut-2 | 284 | 13 | 3.692 | 5 | 7.5 | 1420 | |
| RI0242 | pupa_gut_3 | pupa_gut-3 | 172 | 13 | 2.236 | 8 | 4.5 | 1376 | |
| RI0243 | pupa_gut_4 | pupa_gut-4 | 229 | 13 | 2.977 | 5 | 7.5 | 1145 |
