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protocol_496.ann
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T1 Action 1549 1554 Store
E1 Action:T1 Acts-on:T2
T2 Reagent 1555 1558 DNA
T3 Temperature 1562 1565 4°C
R1 Setting Arg1:E1 Arg2:T3
T4 Action 1398 1406 Incubate
E2 Action:T4
T5 Action 1364 1367 Add
E3 Action:T5 Acts-on:T7 Site:T8
T6 Amount 1368 1372 50µl
T7 Reagent 1373 1382 TE Buffer
T8 Reagent 1390 1396 pellet
R2 Measure Arg1:T7 Arg2:T6
T9 Temperature 1410 1426 room temperature
T10 Modifier 1431 1439 at least
T11 Time 1440 1450 15 minutes
R3 Setting Arg1:E2 Arg2:T9
R4 Mod-Link Arg1:T11 Arg2:T10
R5 Setting Arg1:E2 Arg2:T11
T12 Action 1177 1187 Centrifuge
E4 Action:T12
T13 Action 1214 1220 Decant
E5 Action:T13 Acts-on:T16
T14 Action 1256 1262 Invert
E6 Action:T14 Acts-on:T15
T15 Device 1267 1271 tube
T16 Reagent 1240 1252 ethanol wash
T17 Time 1192 1200 1 minute
T18 Speed 1204 1212 14,000xg
T19 Amount 1098 1103 700µl
T20 Action 1094 1097 Add
E7 Action:T20 Acts-on:T24 Site:T25
T21 Action 1070 1076 Remove
E8 Action:T21
T22 Reagent 1081 1092 supernatant
T23 Generic-Measure 1104 1107 70%
T24 Reagent 1108 1115 ethanol
T25 Location 1123 1127 tube
T26 Action 1132 1138 invert
E9 Action:T26
T27 Generic-Measure 1139 1152 several times
R6 Setting Arg1:E9 Arg2:T27
T28 Modifier 1277 1282 clean
T29 Location 1283 1300 absorbent surface
T30 Time 1305 1320 several minutes
R7 Setting Arg1:E6 Arg2:T30
R8 Mod-Link Arg1:T29 Arg2:T28
R9 Setting Arg1:E4 Arg2:T18
R10 Setting Arg1:E4 Arg2:T17
R11 Measure Arg1:T24 Arg2:T23
R12 Measure Arg1:T24 Arg2:T19
T31 Action 1010 1020 Centrifuge
E10 Action:T31
T32 Speed 1024 1032 14,000xg
T33 Time 1037 1046 5 minutes
R13 Setting Arg1:E10 Arg2:T32
R14 Setting Arg1:E10 Arg2:T33
T34 Action 880 886 Invert
E11 Action:T34 Acts-on:T35
T35 Device 891 896 tubes
T36 Generic-Measure 897 905 10 times
R15 Setting Arg1:E11 Arg2:T36
T37 Action 783 791 Transfer
E12 Action:T37 Acts-on:T38 Site:T39
T38 Reagent 796 807 supernatant
T39 Location 819 823 tube
T40 Modifier 813 818 clean
R16 Mod-Link Arg1:T39 Arg2:T40
T41 Action 828 839 precipitate
E13 Action:T41 Acts-on:T42 Using:T43
T42 Reagent 844 855 genomic DNA
T43 Reagent 867 878 isopropanol
T44 Amount 861 866 500µl
R17 Measure Arg1:T43 Arg2:T44
T45 Action 734 744 Centrifuge
E14 Action:T45 Acts-on:T46
T46 Reagent 749 755 sample
T47 Speed 759 767 14,000xg
T48 Time 772 781 5 minutes
R18 Setting Arg1:E14 Arg2:T47
R19 Setting Arg1:E14 Arg2:T48
T49 Action 656 659 Add
E15 Action:T49 Acts-on:T51
T50 Amount 660 665 100µl
T51 Reagent 666 688 Precipitation Solution
T52 Action 693 696 mix
E16 Action:T52 Using:E17
T54 Device 714 718 tube
T55 Generic-Measure 719 732 several times
T53 Action 700 709 inverting
E17 Action:T53 Acts-on:T54
R20 Setting Arg1:E17 Arg2:T55
R21 Measure Arg1:T51 Arg2:T50
T56 Action 610 618 Incubate
E18 Action:T56 Acts-on:T57
T57 Reagent 623 629 sample
T58 Time 634 646 5-10 minutes
T59 Temperature 650 654 60°C
R22 Setting Arg1:E18 Arg2:T58
R23 Setting Arg1:E18 Arg2:T59
T60 Action 531 534 Add
E19 Action:T60 Acts-on:T62 Site:T63
T61 Amount 535 539 50µl
T62 Reagent 540 562 DNA Stripping Solution
T63 Reagent 570 576 sample
T64 Action 581 587 invert
E20 Action:T64
T65 Generic-Measure 588 601 several times
R24 Measure Arg1:T62 Arg2:T61
R25 Setting Arg1:E20 Arg2:T65
T66 Action 423 433 Centrifuge
E21 Action:T66
T67 Time 438 448 10 minutes
T68 Speed 452 460 14,000xg
T69 Action 475 481 remove
E22 Action:T69 Acts-on:T71
T70 Modifier 465 474 carefully
T71 Seal 486 497 upper phase
R26 Setting Arg1:E21 Arg2:T67
R27 Setting Arg1:E21 Arg2:T68
R28 Mod-Link Arg1:E22 Arg2:T70
T72 Action 357 360 Add
E23 Action:T72 Acts-on:T74
T73 Amount 361 366 200µl
T74 Reagent 367 377 chloroform
T75 Action 382 385 mix
E24 Action:T75 Using:E25
T76 Action 389 398 inverting
E25 Action:T76 Acts-on:T77
T77 Device 403 407 tube
T78 Generic-Measure 408 421 several times
R29 Measure Arg1:T74 Arg2:T73
R30 Setting Arg1:E25 Arg2:T78
T79 Action 311 316 Allow
E26 Action:T79 Acts-on:T81 Using:E27
T80 Action 331 335 cool
E27 Action:T80
T81 Reagent 321 327 sample
T82 Temperature 339 355 room temperature
R31 Setting Arg1:E27 Arg2:T82
T83 Action 269 275 Invert
E28 Action:T83 Acts-on:T84
T84 Device 280 284 tube
T85 Modifier 285 297 periodically
R32 Mod-Link Arg1:E28 Arg2:T85
T86 Action 189 192 Add
E29 Action:T86 Acts-on:T92
T87 Action 73 76 Add
E30 Action:T87 Acts-on:T96 Site:T97 Using:T100
T88 Size 77 84 0.5-1cm
T89 Amount 100 108 50-100mg
T90 Modifier 86 99 approximately
R33 Mod-Link Arg1:T89 Arg2:T90
T91 Amount 193 197 10µl
T92 Reagent 198 219 Proteinase K solution
R34 Measure Arg1:T92 Arg2:T91
T93 Action 224 232 incubate
E31 Action:T93
T94 Temperature 236 240 60°C
T95 Time 245 254 3-4 hours
R35 Setting Arg1:E31 Arg2:T94
R36 Setting Arg1:E31 Arg2:T95
T96 Reagent 110 120 mouse tail
T97 Location 135 155 microcentrifuge tube
T98 Amount 129 134 1.5ml
R37 Measure Arg1:T97 Arg2:T98
R38 Measure Arg1:T96 Arg2:T89
R39 Measure Arg1:T96 Arg2:T88
T99 Amount 161 166 500µl
T100 Reagent 167 187 Genomic Lysis Buffer
R40 Measure Arg1:T100 Arg2:T99