Setup Notes
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Normalize plate reader with Buffer AE for all plates
Quantify all plates
Check for samples above 150 ng/ul
Transfer 20 ul from all wells of a plate needing normalization to new VWR plate
Add 20 ul TE to all wells above 150 ng/ul
Restriction Digestion
(Keep MM and reaction plates on ice)
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Template | EcoR1 MID plate | Pool Plate | Library | |||||
|---|---|---|---|---|---|---|---|---|
WRosenthal_Trout_plate9plate12 | 1 | 2Trout_Pool1-4 | 12TroutRedoPool1 | 1and2 | ||||
WRosenthal_Trout_plate10plate16 | 2 | 2Trout_Pool1-4 | 12TroutRedoPool1 | 1and2 | ||||
WRosenthal_Trout_plate11plate18 | 3 | 2Trout_Pool5-8 | 1 | |||||
WRosenthal_Trout_plate12 | 4 | 2Trout_Pool5-8 | 1 | |||||
WRosenthal_Trout_plate13 | 5 | 2Trout_Pool9-12 | 2 | |||||
WRosenthal_Trout_plate14 | 6 | 2Trout_Pool9-12 | 2 | |||||
WRosenthal_Trout_plate15 | 7 | 2Trout_Pool13-16 | 2 | |||||
WRosenthal_Trout_plate16 | 8 | 2Trout_Pool13-16 | 2 | |||||
WRosenthal_Trout_plate17 | 1 | 2Trout_Pool21-24 | 4 | |||||
WRosenthal_Trout_plate18 | 2 | 2Trout_Pool21-24 | 4 | |||||
WRosenthal_Trout_plate19 | 3 | 2Trout_Pool25-28 | 4 | |||||
WRosenthal_Trout_plate20 | 4 | 2Trout_Pool25-28 | 4 | |||||
WRosenthal_Trout_plate21 | 5 | 2Trout_Pool17-20 | 3 | |||||
WRosenthal_Trout_plate22 | 6 | 2Trout_Pool17-20 | 3 | |||||
WRosenthal_Trout_plate24 | 7 | 2Trout_Pool53-56 | 3 | |||||
WRosenthal_Trout_plate25 | 8 | 2Trout_Pool53-56 | 3 | |||||
MT_YCT_RangewideRADs_plate01 | 1 | 2Trout_Pool29-32 | 52TroutRedoPool2 | 1and2 | ||||
Wrosenthal_PM_YCT_plate2 | 4 | 2TroutRedoPool2 | 1and2 | |||||
MT_YCT_RangewideRADs_plate01 | 5 | 2TroutRedoPool3 | 3and4 | |||||
Wrosenthal_GY21_YCT_tray6 | 6 | 2TroutRedoPool3 | 3and4 | |||||
WY_YCT_2021_plate14 | 7 | 2TroutRedoPool4 | 3and4 | |||||
Wrosenthal_PM_YCT_plate1 | 8 | 2TroutRedoPool4 | 3and4 | |||||
WRosenthal_Trout_plate9 | 1 | 2TroutRedoPool5 | 5and6 | |||||
WRosenthal_Trout_plate14 | 2 | 2TroutRedoPool5 | 5and6 | |||||
WRosenthal_Trout_plate20 | 3 | 2TroutRedoPool6 | 5and6 | |||||
WRosenthal_Trout_plate21 | 4 | 2TroutRedoPool6 | 5and6 | |||||
WY_YCT_2021_plate10 | 5 | 2TroutRedoPool7 | 7and8 | |||||
WY_YCT_2021_plate13 | 6 | 2TroutRedoPool7 | 7and8 | |||||
WRosenthal_Trout_plate10 | 1 | 2TroutRedoPool8 | 7and8 | |||||
WRosenthal_Trout_plate13 | 2 | 2TroutRedoPool8 | 7and8 | |||||
WRosenthal_Trout_plate11 | 1 | 2TroutRedoPool9 | 10and14 | |||||
MT_YCT_RangewideRADs_plate03 | 2 | 2Trout_Pool29-32 | 5 | WY_YCT_2021_plate04 | 3 | 2Trout_Pool33-36 | 52TroutRedoPool9 | 9and12 |
WY_YCT_2021_plate09plate11 | 4 | 2Trout_Pool33-36 | 5 | |||||
WY_YCT_2021_plate10 | 5 | 2Trout_Pool37-40 | 6 | |||||
WY_YCT_2021_plate11 | 6 | 2Trout_Pool37-40 | 66 | 2TroutRedoPool9 | 9and12 | |||
MT_WCT_plate15 | ||||||||
WRosenthal_Trout_plate19 | 3 | 2TroutRedoPool10 | 10and14 | |||||
WY_YCT_2021_plate13plate04 | 7 | 2Trout_Pool49-52 | 62TroutRedoPool11 | 11and13 | ||||
WY_YCT_2021_plate14plate09 | 8 | 2Trout_Pool49-52 | 65 | 2TroutRedoPool11 | 11and13 | |||
Wrosenthal_GY21_YCT_tray1tray3 | 4 | 2Trout_Pool45-48 | 87 | 2TroutRedoPool12 | 9and12 | |||
Wrosenthal_GY21_YCT_tray3tray7 | 5 | 2Trout_Pool45-48 | 8 | |||||
Wrosenthal_GY21_YCT_tray4 | 6 | 2Trout_Pool61-64 | 8 | |||||
Wrosenthal_GY21_YCT_tray6 | 7 | 2Trout_Pool57-60 | 7 | |||||
Wrosenthal_PM_YCT_plate2 | 3 | 2Trout_Pool41-44 | 7 | |||||
Wrosenthal_PM_YCT_plate1 | 2 | 2Trout_Pool41-44 | 7 | |||||
MT_WCT_plate15 | 1 | 2Trout_Pool61-64 | 88 | 2TroutRedoPool12 | 9and12 | |||
WRosenthal_Trout_plate17 | 3 | 2TroutRedoPool13 | 11and13 | |||||
WRosenthal_Trout_plate24 | 4 | 2TroutRedoPool13 | 11and13 | |||||
WRosenthal_Trout_plate15 | 2 | 2TroutRedoPool14 | 10and14 | |||||
WRosenthal_Trout_plate22 | 4 | 2TroutRedoPool14 | 10and14 | |||||
WRosenthal_Trout_plate25 | 8 | 2TroutRedoPool15 | 15and16 | |||||
Wrosenthal_GY21_YCT_tray7 | 8 | 2Trout_Pool57-60tray4 | 6 | 2TroutRedoPool15 | 15and16 | |||
Wrosenthal_GY21_YCT_tray1 | 7 | 2TroutRedoPool16 | 15and16 | |||||
Label reaction plates with MID plate used.
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Reagent | ul/rxn | rxns | ul needed (x 1.6) | |
5x Iproof buffer | 0.425 | 1152 | 784 | 163.2278 |
10 mM dNTPs | 0.4 | 1152 | 738 | 153.6262 |
Primers | 1.33 | 1152 | 2452 | 511869 |
Total | 2.155 | 1152 | 3973.82828 |
Then continue with the last four unlisted steps for GBS1 from above:
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