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protocol_51.ann
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T1 Amount 102 110 10-30mm2
T2 Location 111 118 section
T3 Reagent 122 138 stained material
T4 Amount 144 147 2ml
T5 Location 148 162 microfuge tube
R2 Meronym Arg1:T2 Arg2:T3
T6 Action 164 167 add
E1 Action:T6 Acts-on:T2 Acts-on2:T8 Acts-on3:T10
T7 Amount 168 173 600µl
T8 Reagent 174 186 Lysis Buffer
R5 Meronym Arg1:T3 Arg2:T5
T9 Amount 191 195 10µl
T10 Reagent 196 208 Proteinase K
T11 Action 210 216 Invert
E2 Action:T11
T12 Action 220 223 mix
E3 Action:T12 Using:E2
T13 Action 225 233 Incubate
E4 Action:T13 Acts-on:T14 Using:T17
T14 Reagent 238 244 sample
T15 Temperature 248 252 65°C
T16 Time 257 264 4 hours
T17 Method 270 289 periodic inversions
R7 Setting Arg1:E4 Arg2:T15
R8 Setting Arg1:E4 Arg2:T16
T18 Action 300 304 cool
E5 Action:T18
T19 Temperature 308 324 room temperature
R9 Setting Arg1:E5 Arg2:T19
T20 Action 326 332 remove
E6 Action:T20 Acts-on:T21
T21 Reagent 337 353 stained material
T22 Action 358 364 remove
E7 Action:T22 Acts-on:T24 Acts-on2:T25 Using:T26
T23 Modifier 365 371 excess
T24 Reagent 372 378 buffer
R10 Mod-Link Arg1:T24 Arg2:T23
T25 Reagent 388 396 material
T26 Device 404 411 pipette
T27 Action 416 422 return
E8 Action:T27 Site:T28
T28 Location 430 434 tube
T29 Action 436 439 Add
E9 Action:T29 Acts-on:T31
T30 Amount 440 445 200µl
T31 Reagent 446 456 chloroform
T32 Action 461 464 mix
E10 Action:T32 Using:E11
T33 Action 468 477 inverting
E11 Action:T33 Acts-on:T34 Count:T35
T34 Location 482 486 tube
T35 Numerical 487 500 several times
T36 Action 502 512 Centrifuge
E12 Action:T36
T37 Time 517 527 10 minutes
T38 Speed 531 539 14,000xg
R12 Setting Arg1:E12 Arg2:T37
R13 Setting Arg1:E12 Arg2:T38
T39 Action 554 560 remove
E13 Action:T39 Acts-on:T43 Site:T41
T40 Modifier 544 553 carefully
T41 Location 588 608 microcentrifuge tube
T42 Modifier 582 587 clean
R14 Mod-Link Arg1:T41 Arg2:T42
R15 Mod-Link Arg1:E13 Arg2:T40
T43 Reagent 565 576 upper phase
T44 Action 610 613 Add
E14 Action:T44 Acts-on:T46 Acts-on2:T47
T45 Amount 614 618 50µl
T46 Reagent 619 641 DNA Stripping Solution
T47 Reagent 649 655 sample
T48 Action 660 666 invert
E15 Action:T48 Count:T49
T49 Numerical 667 674 several
T50 Action 684 687 mix
E16 Action:T50 Using:E15
T51 Action 689 697 Incubate
E17 Action:T51 Acts-on:T52
T52 Reagent 702 708 sample
T53 Time 713 725 5-10 minutes
T54 Temperature 729 733 60°C
T55 Action 735 738 Add
E18 Action:T55 Acts-on:T57
R17 Setting Arg1:E17 Arg2:T53
R18 Setting Arg1:E17 Arg2:T54
T56 Amount 739 744 100µl
T57 Reagent 745 767 Precipitation Solution
T58 Action 772 775 mix
E19 Action:T58 Using:E20
T59 Action 779 788 inverting
E20 Action:T59 Acts-on:T60 Count:T61
T60 Location 793 797 tube
T61 Numerical 798 805 several
T62 Action 813 823 Centrifuge
E21 Action:T62 Acts-on:T63
T63 Reagent 828 834 sample
T64 Speed 838 846 14,000xg
T65 Time 851 860 5 minutes
R20 Setting Arg1:E21 Arg2:T64
R21 Setting Arg1:E21 Arg2:T65
T66 Action 862 870 Transfer
E22 Action:T66 Acts-on:T67 Site:T68
T67 Reagent 875 886 supernatant
T68 Location 898 902 tube
T69 Modifier 892 897 clean
R22 Mod-Link Arg1:T68 Arg2:T69
T70 Action 907 918 precipitate
E23 Action:T70 Acts-on:T71 Acts-on2:T73
T71 Reagent 923 934 genomic DNA
T72 Amount 940 945 500µl
T73 Reagent 946 957 isopropanol
T74 Action 959 965 Invert
E24 Action:T74 Acts-on:T75 Count:T76
T75 Location 970 975 tubes
T76 Numerical 976 978 10
T77 Action 988 999 precipitate
E25 Action:T77 Acts-on:T78 Using:E24
T78 Reagent 1004 1007 DNA
T79 Action 1047 1050 add
E26 Action:T79 Acts-on:T81
T80 Amount 1051 1054 2µl
T81 Reagent 1055 1070 Mussel Glycogen
T82 Action 1089 1099 Centrifuge
E27 Action:T82
T83 Speed 1103 1111 14,000xg
T84 Time 1116 1125 5 minutes
T85 Action 1129 1135 pellet
E28 Action:T85 Acts-on:T86 Using:E27
T86 Reagent 1136 1147 genomic DNA
R25 Setting Arg1:E27 Arg2:T83
R26 Setting Arg1:E27 Arg2:T84
T87 Action 1149 1155 Remove
E29 Action:T87 Acts-on:T88
T88 Reagent 1160 1171 supernatant
T89 Action 1173 1176 Add
E30 Action:T89 Acts-on:T92 Site:T93
T90 Amount 1177 1182 700µl
T91 Concentration 1183 1186 70%
T92 Reagent 1187 1194 ethanol
T93 Location 1202 1206 tube
T94 Action 1211 1217 invert
E31 Action:T94 Count:T95
T95 Numerical 1218 1225 several
T96 Action 1235 1239 wash
E32 Action:T96 Acts-on:T97 Using:E31
T97 Reagent 1244 1254 DNA pellet
T98 Action 1256 1266 Centrifuge
E33 Action:T98
T99 Time 1271 1279 1 minute
T100 Speed 1283 1291 14,000xg
R29 Setting Arg1:E33 Arg2:T99
R30 Setting Arg1:E33 Arg2:T100
T101 Action 1293 1299 Decant
E34 Action:T101 Acts-on:T103
T102 Action 1303 1310 pipette
E35 Action:T102 Acts-on:T103
T103 Reagent 1319 1331 ethanol wash
T104 Action 1333 1339 Invert
E36 Action:T104 Acts-on:T105 Site:T107
T105 Location 1344 1348 tube
T106 Modifier 1354 1369 clean absorbent
T107 Location 1370 1377 surface
R31 Mod-Link Arg1:T107 Arg2:T106
T108 Time 1382 1397 several minutes
R32 Setting Arg1:E36 Arg2:T108
T109 Action 1429 1439 drain away
E37 Action:T109 Acts-on:T110 Using:E36
T110 Reagent 1418 1425 ethanol
T111 Modifier 1411 1417 excess
R33 Mod-Link Arg1:T110 Arg2:T111
T112 Action 1441 1444 Add
E38 Action:T112 Acts-on:T114 Acts-on2:T115
T113 Amount 1445 1449 50µl
T114 Reagent 1450 1459 TE Buffer
T115 Reagent 1467 1473 pellet
T116 Action 1475 1483 Incubate
E39 Action:T116
T117 Temperature 1487 1503 room temperature
T118 Time 1517 1527 15 minutes
R35 Setting Arg1:E39 Arg2:T117
R36 Setting Arg1:E39 Arg2:T118
T119 Action 1531 1540 rehydrate
E40 Action:T119 Using:E39
T120 Amount 1552 1555 1µl
T121 Reagent 1556 1571 LongLife™ RNase
T122 Amount 1582 1587 100µl
T123 Reagent 1588 1597 TE Buffer
T124 Action 1605 1610 added
E41 Action:T124 Acts-on:T121 Acts-on2:T123
T125 Action 1626 1631 Store
E42 Action:T125 Acts-on:T126
T126 Reagent 1632 1635 DNA
T127 Temperature 1639 1642 4°C
R39 Setting Arg1:E42 Arg2:T127
T128 Action 1666 1671 store
E43 Action:T128
T129 Temperature 1675 1680 -20°C
T130 Temperature 1684 1689 -80°C
R40 Setting Arg1:E43 Arg2:T129
R1 Measure Arg1:T2 Arg2:T1
R3 Measure Arg1:T5 Arg2:T4
R4 Measure Arg1:T8 Arg2:T7
R6 Measure Arg1:T10 Arg2:T9
R11 Measure Arg1:T31 Arg2:T30
R16 Measure Arg1:T46 Arg2:T45
R19 Measure Arg1:T57 Arg2:T56
R23 Measure Arg1:T73 Arg2:T72
R24 Measure Arg1:T81 Arg2:T80
R27 Measure Arg1:T92 Arg2:T91
R28 Measure Arg1:T92 Arg2:T90
R34 Measure Arg1:T114 Arg2:T113
R37 Measure Arg1:T121 Arg2:T120
R38 Measure Arg1:T123 Arg2:T122
R41 Or Arg1:T129 Arg2:T130