20200205-RT2_geoduck_hemolymph
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SampleSexStage2_tubes?
RNA_tube1_ng.ul
tube1_ul
tube1_Total_yield
RNA_tube2_ng.ul
tube2_ul
tube2_Total_yield
Total_yield
cDNA_ul
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19F2Y4.6418.435.813465.4483.8
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21F2N1813234234
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31F4N25994248502485015
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37F6N1344111478414784
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39F5N79917189718915
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55(2)F3N5713741741
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57F7N20881760176015
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61F7N47.7904293429315
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27M2N1341317421742
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28M1N18.713243.1243.1
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43M4Y3.621347.061651219802027.06
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54M3N111.9121342.81342.8
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59M4N1001313001300
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55FF31351216201620
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DNase
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SampleRNA_(ng/ul)RNA.ul
H20_to_44ul
10x Turbo DNase Buffer.ul
Turbo DNase.ul
Total_Rxn_vol
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1935.813.031.05150.0
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211813.031.05150.0
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2818.713.031.05150.0
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3713441.542.55150.0
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43A16512.032.05150.0
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43B3.6213.031.05150.0
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54111.912.032.05150.0
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55F13512.032.05150.0
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55(2)5713.031.05150.0
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Reverse Transcriptase
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SampleRNA_(ng/ul)vol_to_100ng(uL)vol_to_36.5ul_H2Ovol_primer(ul)1:1000_dilution_vol_primerTotal_volNotes
Pooled Sample
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197.9212.6318.870.005536.50
SexStage
Pooled sample
RNA (ng/ul)
ul_RNA
Total_yield(ng)
Pooled_ul
Pooled_ng
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213.7226.884.620.005536.50F66147.79042932.095.4
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3739.62.5328.970.005536.50F355F27.645124221.0579.6
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55(2)14.56.9024.600.005536.50M443B0.5964526.8245.026.82
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271340.7530.750.005536.50F2194.6418.44.018.4
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283.5728.013.490.005536.5072.0720.22
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4336.82.7228.780.005536.50
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5424.44.1027.400.005536.50
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591001.0030.500.005536.50
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31254.0027.500.005536.50
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39791.2730.230.005536.50
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57205.0026.500.005536.50
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6147.72.1029.400.005536.50
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TotalRNA_ng
Total_ulVol_to_73For_4X_RXN1:10_dilution_vol_primer
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pooled720.2272.00.00.1173.0
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Master MixSingle Rxn Vol#RxnsTotal
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Buffer 532160
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dNTPs1.253240
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M-MLV RT0.025320.8
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qPCR
60C_anneal;4ul_template
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For each primer (17):
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Master Mix
Final Concentration
Single Rxn vol
#RxnsTotal
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2x SsoFast EvaGreen Supermix
1x10330
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Fwd Primer0.2um0.531.5
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Rvs Primer0.2um0.531.5
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Water
bring rxn vol to 20ul
Variable315.00
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