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{"sheet":"Sheet1","index":0,"n_rows":31,"n_cols":56,"rows_kept":31,"cut":false,"hidden":false,"merged":[[23,7,30,7],[23,8,30,8],[21,2,21,3],[21,4,21,7],[23,4,30,4],[23,5,30,5]]}
[{"$nulls":14},"WT averaged fluorescence",{"$nulls":13},"TAC-averaged signal"]
[null,"Raw Fluorescence data",1,2,3,4,5,6,7,8,9,10,11,12,{"$f":"AVERAGE(C3:H3)","v":536.16666666666663},"Background-normalized incorporation data",1,2,3,4,5,6,7,8,9,10,11,12,{"$f":"AVERAGE(AE4:AJ4)","v":65757.49093139614},"OD-normalized incorporation data",1,2,3,4,5,6,7,8,9,10,11,12,null,"Final data",1,2,3,4,5,6,7,8,9,10,11,12]
[null,"A",546,633,568,518,519,433,1858,1872,1730,1959,1991,1867,"Read 1:488,530","A",{"$f":"C3-0.9*$O$2","v":63.450000000000045},150.45000000000005,85.450000000000045,35.450000000000045,36.450000000000045,-49.549999999999955,1375.45,1389.45,1247.45,1476.45,1508.45,1384.45,null,"A",{"$f":"Q3/Q13","v":755.35714285714346},{"$f":"R3/R13","v":1791.0714285714294},1005.2941176470595,322.27272727272771,340.65420560747714,-481.06796116504819,16980.864197530867,17813.461538461543,15593.125000000004,15541.578947368422,16396.195652173912,15382.777777777779,null,"A",{"$f":"AE3/$AC$2","v":1.1487012843072082E-2},{"$f":"AF3/$AC$2","v":2.7237526906858853E-2},1.528790261623377E-2,4.9009279811018077E-3,5.1804623440221713E-3,-7.3157894918305134E-3,0.25823467344955059,0.27089630833156447,0.23713077824499251,0.23634689717073804,0.24934338917036586,0.23393194539351286]
[null,"B",7693,7714,7822,6377,7152,7875,841,843,887,540,567,560,"Read 1:488,530","B",7210.45,7231.45,7339.45,5894.45,6669.45,7392.45,358.45000000000005,360.45000000000005,404.45000000000005,57.450000000000045,84.450000000000045,77.450000000000045,null,"B",{"$f":"Q4/Q14","v":67387.383177570096},{"$f":"R4/R14","v":66957.87037037038},{"$f":"S4/S14","v":62730.341880341883},{"$f":"T4/T14","v":60147.448979591834},{"$f":"U4/U14","v":64129.326923076929},{"$f":"V4/V14","v":73192.57425742576},{"$f":"W4/W14","v":3350.0000000000009},{"$f":"X4/X14","v":3276.8181818181824},{"$f":"Y4/Y14","v":3516.9565217391309},{"$f":"Z4/Z14","v":541.98113207547226},{"$f":"AA4/AA14","v":812.0192307692314},{"$f":"AB4/AB14","v":766.83168316831745},null,"B",{"$f":"AE4/$AC$2","v":1.0247864117545848},{"$f":"AF4/$AC$2","v":1.0182546417445668},{"$f":"AG4/$AC$2","v":0.95396495504652934},{"$f":"AH4/$AC$2","v":0.91468588791416661},{"$f":"AI4/$AC$2","v":0.97523987023747838},{"$f":"AJ4/$AC$2","v":1.1130682333026747},{"$f":"AK4/$AC$2","v":5.0944766178731005E-2},{"$f":"AL4/$AC$2","v":4.9831861517296037E-2},{"$f":"AM4/$AC$2","v":5.348373960022778E-2},{"$f":"AN4/$AC$2","v":8.242120013990703E-3},{"$f":"AO4/$AC$2","v":1.2348695475872088E-2},{"$f":"AP4/$AC$2","v":1.1661510685806764E-2}]
[null,"C",5898,5926,6326,704,677,758,1395,1366,1486,664,642,724,"Read 1:488,530","C",5415.45,5443.45,5843.45,221.45000000000005,194.45000000000005,275.45000000000005,912.45,883.45,1003.45,181.45000000000005,159.45000000000005,241.45000000000005,null,"C",58230.645161290318,49939.908256880735,52643.693693693698,2171.0784313725499,1984.1836734693882,2899.4736842105267,11127.439024390245,11043.125000000002,12701.898734177217,1814.5000000000005,1752.1978021978027,2541.5789473684217,null,"C",0.88553629915778764,0.7594558057116616,0.80057333313730172,3.3016442699092721E-2,3.0174260686732391E-2,4.4093435487608655E-2,0.16921933709422315,0.16793714059925335,0.19316276448912761,2.7593814397405197E-2,2.6646360397568177E-2,3.8650789611483431E-2]
[null,"D",6152,6263,6419,937,913,1032,646,628,608,539,597,720,"Read 1:488,530","D",5669.45,5780.45,5936.45,454.45000000000005,430.45000000000005,549.45000000000005,163.45000000000005,145.45000000000005,125.45000000000005,56.450000000000045,114.45000000000005,237.45000000000005,null,"D",27928.32512315271,28060.436893203885,28678.502415458937,4783.6842105263158,4628.4946236559144,6105.0000000000009,1969.2771084337357,1598.3516483516489,1441.9540229885063,424.43609022556433,1244.0217391304352,2638.3333333333339,null,"D",0.4247170128843562,0.42672608847680854,0.43612525370499328,7.2747365247209109E-2,7.0387336227362401E-2,9.2841133588403804E-2,2.9947570695607207E-2,2.4306761491541498E-2,2.192836135571043E-2,6.4545663804048197E-3,1.8918327349629345E-2,4.0122171572602575E-2]
[null,"E",4411,4617,4520,780,785,785,744,883,894,671,691,674,"Read 1:488,530","E",3928.45,4134.45,4037.45,297.45000000000005,302.45000000000005,302.45000000000005,261.45000000000005,400.45000000000005,411.45000000000005,188.45000000000005,208.45000000000005,191.45000000000005,null,"E",48499.382716049389,52334.810126582284,52434.415584415576,3198.3870967741941,3252.1505376344089,3516.86046511628,3395.4545454545455,5133.9743589743603,5143.1250000000009,2141.4772727272734,2127.0408163265311,2103.8461538461543,null,"E",0.7375491678453503,0.79587601937522912,0.79739075870640597,4.863912919230793E-2,4.9456730960543059E-2,5.3482278829424482E-2,5.1636011310057059E-2,7.8074365159865405E-2,7.8213522553130119E-2,3.2566286249599251E-2,3.2346745385185741E-2,3.1994015039914878E-2]
[null,"F",759,808,813,675,672,667,767,801,767,622,684,685,"Read 1:488,530","F",276.45000000000005,325.45000000000005,330.45000000000005,192.45000000000005,189.45000000000005,184.45000000000005,284.45000000000005,318.45000000000005,284.45000000000005,139.45000000000005,201.45000000000005,202.45000000000005,null,"F",3037.9120879120883,3740.8045977011502,3755.1136363636369,2004.6875000000005,2015.425531914894,1882.1428571428576,25859.090909090923,9951.5625000000036,7901.3888888888905,7339.4736842105303,15496.153846153853,15573.076923076929,null,"F",4.6198722683648302E-2,5.6887885238867744E-2,5.7105488411674558E-2,3.0486070432514868E-2,3.0649367902701136E-2,2.8622485902122855E-2,0.39324935521139859,0.15133732079866502,0.12015952520348287,0.11161425991553874,0.23565609980953761,0.23682589926255093]
[null,"G",4207,3897,4291,843,735,810,1772,1728,1973,911,877,940,"Read 1:488,530","G",3724.45,3414.45,3808.45,360.45000000000005,252.45000000000005,327.45000000000005,1289.45,1245.45,1490.45,428.45000000000005,394.45000000000005,457.45000000000005,null,"G",30528.278688524588,29183.333333333332,31216.803278688523,3499.5145631067971,2450.9708737864084,3179.1262135922339,10315.6,10378.75,11465,4327.7777777777783,4241.3978494623661,4972.2826086956529,null,"G",0.46425552824657362,0.44380241581571128,0.47472619220305368,5.3218492882549172E-2,3.7272877037590621E-2,4.8346221374367393E-2,0.15687338208755744,0.15783372894850875,0.17435275947437337,6.5814217003700576E-2,6.4500603496069467E-2,7.5615455186438987E-2]
[null,"H",4786,4455,4426,740,706,711,492,435,442,463,402,415,"Read 1:488,530","H",4303.45,3972.45,3943.45,257.45000000000005,223.45000000000005,228.45000000000005,9.4500000000000455,-47.549999999999955,-40.549999999999955,-19.549999999999955,-80.549999999999955,-67.549999999999955,null,"H",47290.659340659338,45660.34482758621,41510,2681.7708333333339,2377.1276595744685,2284.5000000000005,{"$nulls":7},"H",0.71916763658146587,0.69437480324824141,0.63125887882958909,4.0782742701226046E-2,3.6149914266870253E-2,3.4741289055316711E-2]
[]
[null,"Raw OD data",1,2,3,4,5,6,7,8,9,10,11,12,null,"Normalized OD data",1,2,3,4,5,6,7,8,9,10,11,12,{"$nulls":17},"Averages",null,null,null,"Std. Deviation"]
[null,"A",0.124,0.124,0.125,0.15,0.14699999999999999,0.14299999999999999,0.121,0.11799999999999999,0.12,0.13500000000000001,0.13200000000000001,0.13,"Read 2:600","A",{"$f":"C13-0.04","v":8.3999999999999991E-2},{"$f":"D13-0.04","v":8.3999999999999991E-2},8.4999999999999992E-2,0.10999999999999999,0.10699999999999998,0.10299999999999998,8.0999999999999989E-2,7.7999999999999986E-2,7.9999999999999988E-2,9.5000000000000001E-2,9.1999999999999998E-2,0.09,{"$nulls":17},"+ nsAA","- nsAA",null,null,"+ nsAA","- nsAA"]
[null,"B",0.14699999999999999,0.14799999999999999,0.157,0.13800000000000001,0.14399999999999999,0.14099999999999999,0.14699999999999999,0.15,0.155,0.14599999999999999,0.14399999999999999,0.14099999999999999,"Read 2:600","B",{"$f":"C14-0.04","v":0.10699999999999998},{"$f":"D14-0.04","v":0.10799999999999998},{"$f":"E14-0.04","v":0.11699999999999999},{"$f":"F14-0.04","v":9.8000000000000004E-2},{"$f":"G14-0.04","v":0.10399999999999998},{"$f":"H14-0.04","v":0.10099999999999998},{"$f":"I14-0.04","v":0.10699999999999998},{"$f":"J14-0.04","v":0.10999999999999999},{"$f":"K14-0.04","v":0.11499999999999999},{"$f":"L14-0.04","v":0.10599999999999998},{"$f":"M14-0.04","v":0.10399999999999998},{"$f":"N14-0.04","v":0.10099999999999998},{"$nulls":16},"WT Py79",{"$f":"AVERAGE(AS3:AU3)","v":1.8004147455388236E-2},{"$f":"AVERAGE(AV3:AX3)","v":9.2186694443115516E-4},null,"WT Py79",{"$f":"_xlfn.STDEV.P(AS3:AU3)","v":6.7108450952201798E-3},{"$f":"_xlfn.STDEV.P(AV3:AX3)","v":5.8260205135481514E-3}]
[null,"C",0.13300000000000001,0.14899999999999999,0.151,0.14199999999999999,0.13800000000000001,0.13500000000000001,0.122,0.12,0.11899999999999999,0.14000000000000001,0.13100000000000001,0.13500000000000001,"Read 2:600","C",9.2999999999999999E-2,0.10899999999999999,0.11099999999999999,0.10199999999999998,9.8000000000000004E-2,9.5000000000000001E-2,8.199999999999999E-2,7.9999999999999988E-2,7.8999999999999987E-2,0.1,9.0999999999999998E-2,9.5000000000000001E-2,{"$nulls":16},"TAC + ctrl",{"$f":"AVERAGE(AS4:AU4)","v":0.99900200284856033},{"$f":"AVERAGE(AV4:AX4)","v":1.0009979971514398},null,"TAC + ctrl",3.1957448384873299E-2,8.3012045579936328E-2]
[null,"D",0.24299999999999999,0.246,0.247,0.13500000000000001,0.13300000000000001,0.13,0.123,0.13100000000000001,0.127,0.17299999999999999,0.13200000000000001,0.13,"Read 2:600","D",0.20299999999999999,0.20599999999999999,0.20699999999999999,9.5000000000000001E-2,9.2999999999999999E-2,0.09,8.299999999999999E-2,9.0999999999999998E-2,8.6999999999999994E-2,0.13299999999999998,9.1999999999999998E-2,0.09,{"$nulls":16},"Strain/nsAA 1",0.81518847933558369,3.5761379624477924E-2,null,"Strain/nsAA 1",5.2499362413287294E-2,6.0048239401109442E-3]
[null,"E",0.121,0.11899999999999999,0.11700000000000001,0.13300000000000001,0.13300000000000001,0.126,0.11700000000000001,0.11799999999999999,0.12,0.128,0.13800000000000001,0.13100000000000001,"Read 2:600","E",8.0999999999999989E-2,7.8999999999999987E-2,7.7000000000000013E-2,9.2999999999999999E-2,9.2999999999999999E-2,8.5999999999999993E-2,7.7000000000000013E-2,7.7999999999999986E-2,7.9999999999999988E-2,8.7999999999999995E-2,9.8000000000000004E-2,9.0999999999999998E-2,{"$nulls":16},"Strain/nsAA 2",0.42918945168871936,7.8658611687658433E-2,null,"Strain/nsAA 2",4.9724647403314405E-3,1.0074733407307303E-2]
[null,"F",0.13100000000000001,0.127,0.128,0.13600000000000001,0.13400000000000001,0.13800000000000001,5.0999999999999997E-2,7.1999999999999995E-2,7.5999999999999998E-2,5.8999999999999997E-2,5.2999999999999999E-2,5.2999999999999999E-2,"Read 2:600","F",9.0999999999999998E-2,8.6999999999999994E-2,8.7999999999999995E-2,9.6000000000000002E-2,9.4E-2,9.8000000000000004E-2,1.0999999999999996E-2,3.1999999999999994E-2,3.5999999999999997E-2,1.8999999999999996E-2,1.2999999999999998E-2,1.2999999999999998E-2,{"$nulls":16},"Strain/nsAA 3",0.77693864864232831,5.0526046327425155E-2,null,"Strain/nsAA 3",2.785943295548789E-2,2.1168531861016711E-3]
[null,"G",0.16200000000000001,0.157,0.16200000000000001,0.14299999999999999,0.14299999999999999,0.14299999999999999,0.16500000000000001,0.16,0.17,0.13900000000000001,0.13300000000000001,0.13200000000000001,"Read 2:600","G",0.122,0.11699999999999999,0.122,0.10299999999999998,0.10299999999999998,0.10299999999999998,0.125,0.12,0.13,9.9000000000000005E-2,9.2999999999999999E-2,9.1999999999999998E-2,{"$nulls":16},"Strain/nsAA 4",5.3397365444730199E-2,2.9919308079112954E-2,null,"Strain/nsAA 4",5.0909842523465948E-3,9.1941188992105587E-4]
[null,"H",0.13100000000000001,0.127,0.13500000000000001,0.13600000000000001,0.13400000000000001,0.14000000000000001,4.1000000000000009E-2,5.7000000000000023E-2,4.1000000000000009E-2,3.8000000000000006E-2,3.9000000000000007E-2,4.200000000000001E-2,"Read 2:600","H",9.0999999999999998E-2,8.6999999999999994E-2,9.5000000000000001E-2,9.6000000000000002E-2,9.4E-2,0.1,1.0000000000000078E-3,1.7000000000000022E-2,1.0000000000000078E-3,-1.9999999999999948E-3,-9.9999999999999395E-4,2.0000000000000087E-3,{"$nulls":16},"Strain/nsAA 5",0.46092804542177951,4.6279197098169057E-2,null,"Strain/nsAA 5",1.2841964880177373E-2,6.6718367379876844E-3]
[null,null,"Suggested plate layout",{"$nulls":41},"Strain/nsAA 6",0.68160043955309879,3.7224648674471006E-2,null,"Strain/nsAA 6",3.7007888332366588E-2,2.5808372122341104E-3]
[{"$nulls":4},"Columns",{"$nulls":5},"Data analysis: ",{"$nulls":33},"Strain/nsAA 7",{"$f":"AVERAGE(AY3:BA3)","v":0.25542058667536921},{"$f":"AVERAGE(BB3:BD3)","v":0.23987407724487228},null,"Strain/nsAA 7",{"$f":"_xlfn.STDEV.P(AY3:BA3)","v":1.3927600147100387E-2},1.6105165848694529E-2]
[null,null,"Rows",null,"1-3","4-6",null,"7-9","10-12",null,"1. Average technical replicates of fluorescence/luminescence data from the WT controls without nsAA. In this case WT is in wells A 1-3 and have an average fluorescence of 536.1667, represented in cell O2.",{"$nulls":33},"Strain/nsAA 8",5.1420122432084943E-2,1.0750775391889851E-2,null,"Strain/nsAA 8",1.5282944870783951E-3,2.0520577625155099E-2]
[null,null,"A","WT Py79","+ nsAA"," -nsAA","Strain/nsAA 7","+ nsAA"," -nsAA",null," then subtract 90% of that value from the fluorescence/luminescence values of all samples. Only 90% of the value is used to avoid negative numbers. This results in background-normalized incorporation data, shown in cells P-AB:2-10",{"$nulls":33},"Strain/nsAA 9",0.17677308072753473,3.0963654802152268E-2,null,"Strain/nsAA 9",1.1601071999796538E-2,7.2835933244600429E-2]
[null,null,"B","TAC + ctrl",null,null,"Strain/nsAA 8",null,null,null,"2. Subtract plate reader background OD (from reading blank media wells, should be constant per given plate reader) from OD values. This can be anywhere between 0.001 - 0.04 for many plate readers. For this plate reader it is known to be",{"$nulls":33},"Strain/nsAA 10",2.5394231180953044E-2,2.1831688434212248E-2,null,"Strain/nsAA 10",3.3629225735277305E-3,7.9147961890654681E-3]
[null,null,"C","Strain/nsAA 1",null,null,"Strain/nsAA 9",null,null,null,"0.04 as seein in wells H7-H12, but you should include several blank media wells until you know this value for your plate reader. This results in normalized OD data present in cells P-AB:12-20. ",{"$nulls":33},"Strain/nsAA 11",6.9307966341017532E-2,3.2302348891566621E-2,null,"Strain/nsAA 11",1.2496088378828596E-2,2.1485588950719461E-2]
[null,null,"D","Strain/nsAA 2",null,null,"Strain/nsAA 10",null,null,null,"Note any especially low OD values and consider data from those wells suspect. Most nsAAs described here should not significantly hinder growth except for 5OHW which was observed to decrease final OD. ",{"$nulls":33},"Strain/nsAA 12",0.22158206707118214,0.19469875299587577,null,"Strain/nsAA 12",0.12205260368001389,1.7071737732745552E-2]
[null,null,"E","Strain/nsAA 3",null,null,"Strain/nsAA 11",null,null,null,"3. Divide background-normalized incorporation data by the normalized OD data. This results in OD-normalized incorporation data. Skip this step for timecourse analysis. ",{"$nulls":33},"Strain/nsAA 13",0.16301995683681317,6.8643425228736343E-2,null,"Strain/nsAA 13",8.0230866146703186E-3,4.7750606918461919E-2]
[null,null,"F","Strain/nsAA 4",null,null,"Strain/nsAA 12",null,null,null,"4. Divide all averages by OD-normalized fluorescence/luminescence data from the TAC positive control lacking a TAG stop codon."]
[null,null,"G","Strain/nsAA 5",null,null,"Strain/nsAA 13",null,null,null," This results in final data as a fraction of protein expression without nsAA incorporation, and is a good way to reduce inter-experiment variation in values. "]
[null,null,"H","Strain/nsAA 6",null,null,"Blank media",null,null,null,"5. Use standard deviation among three biological replicates to plot error bars. "]