Pooled 6 new samples for sequencing

Today I pooled samples for the 6 new libraries for sequencing. There is over 1000ng in each pooled sample, so everything is ready to go! Details in post.

Pooling

I pooled 110ng of RNA from 10 samples within each of the 6 libraries. Math below:

FRP pool_number trtmnt_tank day infection_status maturity tube_number total-yield_ng elution_vol vol_for_pool
6112 5 NA 9 0 I 28 287.3 13 4.977
6136 5 NA 9 0 M 91 204.1 13 7.006
6143 5 NA 9 0 M 10 297.7 13 4.803
6150 5 NA 9 0 M 119 419.9 13 3.406
6161 5 NA 9 0 M 53 405.6 13 3.526
6213 5 NA 9 0 I 108 176.8 13 8.088
6106 5 NA 9 0 M 5 174.2 13 8.209
6156 5 NA 9 0 I 24 366.6 13 3.901
6157 5 NA 9 0 M 169 365.3 13 3.915
6178 5 NA 9 0 M 124 296.4 13 4.825
                   
6122 6 NA 9 1 I 133 305.5 13 4.681
6201 6 NA 9 1 I 163 211.9 13 6.748
6206 6 NA 9 1 I 103 208 13 6.875
6174 6 NA 9 1 I 20 360.1 13 3.971
6177 6 NA 9 1 I 54 327.6 13 4.365
6187 6 NA 9 1 I 158 241.8 13 5.914
6199 6 NA 9 1 I 151 241.8 13 5.914
6231 6 NA 9 1 I 102 188.5 13 7.586
6233 6 NA 9 1 I 117 300.3 13 4.762
6237 6 NA 9 1 I 17 226.2 13 6.322
                   
6104 7 cold 12 0 I 259 412.1 13 3.470
6106 7 cold 12 0 M 241 169 13 8.462
6153 7 cold 12 0 M 209 315.9 13 4.527
6157 7 cold 12 0 M 224 249.6 13 5.729
6160 7 cold 12 0 M 238 494 13 2.895
6172 7 cold 12 0 M 316 373.1 13 3.833
6175 7 cold 12 0 I 246 269.1 13 5.314
6178 7 cold 12 0 M 218 162.5 13 8.800
6189 7 cold 12 0 I 216 577.2 13 2.477
6191 7 cold 12 0 I 227 234 13 6.111
                   
6118 8 cold 12 1 I 240 280.8 13 5.093
6128 8 cold 12 1 I 231 492.7 13 2.902
6141 8 cold 12 1 I 249 241.8 13 5.914
6148 8 cold 12 1 I 213 442 13 3.235
6149 8 cold 12 1 I 226 340.6 13 4.198
6163 8 cold 12 1 I 228 364 13 3.929
6164 8 cold 12 1 I 257 353.6 13 4.044
6173 8 cold 12 1 I 210 432.9 13 3.303
6174 8 cold 12 1 I 233 457.6 13 3.125
6177 8 cold 12 1 I 250 410.8 13 3.481
                   
6223 9 warm 12 0 M 270 380.9 13 3.754
6228 9 warm 12 0 I 290 370.5 13 3.860
6230 9 warm 12 0 M 292 223.6 13 6.395
6232 9 warm 12 0 M 286 663 13 2.157
6234 9 warm 12 0 I 263 345.8 13 4.135
6235 9 warm 12 0 I 297 531.7 13 2.689
6238 9 warm 12 0 M 375 301.6 13 4.741
6242 9 warm 12 0 M 377 289.9 13 4.933
6253 9 warm 12 0 I 275 387.4 13 3.691
6265 9 warm 12 0 I 282 317.2 13 4.508
                   
6231 10 warm 12 1 I 278 230.1 13 6.215
6233 10 warm 12 1 I 371 370.5 13 3.860
6244 10 warm 12 1 I 284 358.8 13 3.986
6245 10 warm 12 1 I 365 481 13 2.973
6249 10 warm 12 1 I 279 461.5 13 3.099
6255 10 warm 12 1 I 262 536.9 13 2.663
6256 10 warm 12 1 I 283 247 13 5.789
6257 10 warm 12 1 I 363 269.1 13 5.314
6261 10 warm 12 1 I 273 469.3 13 3.047
6262 10 warm 12 1 I 289 352.3 13 4.059

Results:

Ran 2ul of each pooled sample on Qubit RNA Hs

trtmnt_tank day infection_status total-yield_ng elution_vol vol_for_pool total_vol_ul qubit_vol_ul qubit_sample_ng total_ng_RNA_pool final_sample_vol
NA 9 0 287.3 13 4.977375566 52.65550411 2 21 1063.765586 50.65550411
NA 9 1 305.5 13 4.680851064 57.13842355 2 21.5 1185.476106 55.13842355
cold 12 0 412.1 13 3.470031546 51.61757335 2 23.6 1170.974731 49.61757335
cold 12 1 280.8 13 5.092592593 39.22471819 2 27.2 1012.512335 37.22471819
warm 12 0 380.9 13 3.754266212 40.86453844 2 26.9 1045.456084 38.86453844
warm 12 1 230.1 13 6.214689266 41.00446376 2 26.3 1025.817397 39.00446376

There’s at least 1000ng of RNA in all pooled samples!! :fireworks: :tada:

Written on November 22, 2019