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protocol_399.ann
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T1 Action 37 44 Prepare
E1 Action:T1 Creates:T28 Acts-on:T29 Using:T30
T2 Action 155 164 resuspend
E2 Action:T2 Acts-on:T31 Site:T32
T3 Action 263 267 Keep
E3 Action:T3 Acts-on:T39 Site:T40 Using:T42
T4 Action 318 324 Filter
E4 Action:T4 Acts-on:T43 Using:T45
T5 Action 363 373 centrifuge
E5 Action:T5
T6 Action 404 413 resuspend
E6 Action:T6 Site:T50
T7 Action 460 465 Count
E7 Action:T7
T8 Action 470 476 adjust
E8 Action:T8
T9 Action 521 528 Aliquot
E9 Action:T9 Acts-on:T54 Site:T56
T10 Action 585 594 Resuspend
E10 Action:T10 Acts-on:T57 Using:E29
T11 Action 645 648 Add
E11 Action:T11 Acts-on:T62
T12 Action 669 672 mix
E12 Action:T12
T13 Action 682 690 incubate
E13 Action:T13 Acts-on:T64
T14 Action 714 722 Scale up
E14 Action:T14
T15 Action 785 788 add
E15 Action:T15 Acts-on:T70
T17 Action 891 894 Add
E17 Action:T17 Acts-on:T16
T18 Action 927 937 centrifuge
E18 Action:T18 Acts-on:T73
T19 Action 974 983 Resuspend
E19 Action:T19 Acts-on:T76 Site:T78
T20 Action 1033 1037 Take
E20 Action:T20 Acts-on:T79
T21 Action 1117 1122 Place
E21 Action:T21 Acts-on:T85 Site:T86
T22 Action 1161 1169 Pour out
E22 Action:T22 Acts-on:T88
T23 Action 1182 1191 Resuspend
E23 Action:T23 Acts-on:T90 Site:T93
T24 Action 1243 1249 Repeat
E24 Action:T24 Using:T94 Acts-on:T96 Count:T97
T25 Action 1349 1353 Take
E25 Action:T25 Acts-on:T99
T26 Action 1411 1415 pool
E26 Action:T26 Acts-on:T103
T27 Action 1444 1451 process
E27 Action:T27 Acts-on:T106
T28 Reagent 45 50 cells
T29 Reagent 56 79 your tissue of interest
T30 Method 80 107 without lysing erythrocytes
T31 Reagent 169 174 cells
T32 Reagent 178 194 MojoSort™ Buffer
T33 Action 198 204 adding
E28 Action:T33 Site:T37 Acts-on:T31
T34 Amount 211 215 4 mL
T35 Amount 221 225 5 mL
T36 Size 227 237 12 x 75 mm
T37 Location 239 255 polystyrene tube
R1 Measure Arg1:T37 Arg2:T36
R2 Measure Arg1:T37 Arg2:T35
T38 Modifier 205 210 up to
R4 Mod-Link Arg1:T34 Arg2:T38
R3 Measure Arg1:T31 Arg2:T34
T39 Reagent 268 284 MojoSort™ Buffer
T40 Location 288 291 ice
T41 Modifier 292 302 throughout
T42 Method 307 316 procedure
T43 Reagent 329 334 cells
T44 Size 342 347 70 μm
T45 Device 348 361 cell strainer
T46 Speed 377 384 300 x g
T47 Time 389 398 5 minutes
R5 Setting Arg1:E5 Arg2:T46
R6 Setting Arg1:E5 Arg2:T47
R7 Setting Arg1:T45 Arg2:T44
T48 Modifier 420 431 appropriate
T49 Amount 432 438 volume
T50 Reagent 442 458 MojoSort™ Buffer
R8 Measure Arg1:T50 Arg2:T49
R9 Mod-Link Arg1:T49 Arg2:T48
T51 Measure-Type 481 499 cell concentration
T52 Generic-Measure 503 519 1 x 108 cells/mL
R10 Or Arg1:T52 Arg2:T51
R11 Setting Arg1:E8 Arg2:T52
R12 Setting Arg1:E7 Arg2:T52
T53 Amount 529 535 100 μL
T54 Reagent 539 554 cell suspension
T55 Modifier 574 577 new
T56 Location 578 582 tube
R13 Mod-Link Arg1:T56 Arg2:T55
R14 Measure Arg1:T54 Arg2:T53
T57 Reagent 599 604 beads
T58 Action 608 617 vortexing
E29 Action:T58
T59 Speed 619 632 maximum speed
T60 Generic-Measure 634 643 5 touches
R15 Setting Arg1:E29 Arg2:T59
R16 Setting Arg1:E29 Arg2:T60
T61 Amount 649 654 10 μL
T62 Reagent 658 667 Nanobeads
T63 Modifier 673 677 well
T64 Location 694 697 ice
T65 Time 702 712 15 minutes
R17 Setting Arg1:E13 Arg2:T65
R18 Mod-Link Arg1:E12 Arg2:T63
R19 Measure Arg1:T62 Arg2:T61
T66 Amount 727 733 volume
T67 Modifier 734 745 accordingly
R20 Setting Arg1:E14 Arg2:T66
R21 Mod-Link Arg1:E14 Arg2:T67
T68 Amount 789 795 100 μL
T69 Generic-Measure 800 807 1 x 108
T70 Reagent 808 813 cells
R22 Measure Arg1:T70 Arg2:T69
R23 Measure Arg1:T70 Arg2:T68
T16 Reagent 895 911 MojoSort™ Buffer
T71 Amount 918 922 4 mL
T72 Modifier 912 917 up to
T73 Reagent 942 947 cells
T74 Speed 951 958 300 x g
T75 Time 963 972 5 minutes
R24 Setting Arg1:E18 Arg2:T74
R25 Setting Arg1:E18 Arg2:T75
R26 Mod-Link Arg1:T71 Arg2:T72
R27 Measure Arg1:T16 Arg2:T71
T76 Reagent 988 993 cells
T77 Amount 997 1001 3 mL
T78 Reagent 1005 1021 MojoSort™ Buffer
R28 Measure Arg1:T78 Arg2:T77
T79 Reagent 1041 1048 aliquot
T80 Location 1068 1072 tube
T81 Device 1080 1086 magnet
T82 Action 1090 1097 monitor
E16 Action:T82 Measure-Type-Link:T83 Using:E30
T83 Measure-Type 1098 1114 purity and yield
T84 Action 1056 1063 placing
E30 Action:T84 Acts-on:T80 Site:T81
T85 Location 1127 1131 tube
T86 Device 1139 1145 magnet
T87 Time 1150 1159 5 minutes
R29 Setting Arg1:E21 Arg2:T87
T88 Reagent 1174 1180 liquid
T89 Modifier 1196 1203 labeled
T90 Reagent 1204 1209 cells
T91 Modifier 1213 1224 appropriate
T92 Amount 1225 1231 amount
T93 Reagent 1235 1241 buffer
R30 Measure Arg1:T93 Arg2:T92
R31 Mod-Link Arg1:T92 Arg2:T91
R32 Mod-Link Arg1:T90 Arg2:T89
T94 Method 1250 1261 steps 7 – 9
T95 Modifier 1269 1276 labeled
T96 Reagent 1277 1285 fraction
T97 Numerical 1286 1287 2
R33 Mod-Link Arg1:T96 Arg2:T95
T98 Modifier 1356 1361 small
T99 Reagent 1362 1369 aliquot
R34 Mod-Link Arg1:T99 Arg2:T98
T100 Action 1373 1380 monitor
E31 Action:T100 Measure-Type-Link:T101 Using:E25
T101 Measure-Type 1381 1397 purity and yield
T102 Modifier 1420 1429 unlabeled
T103 Reagent 1430 1439 fractions
R35 Mod-Link Arg1:T103 Arg2:T102
T104 Modifier 1452 1466 simultaneously
R36 Mod-Link Arg1:E27 Arg2:T104
T105 Modifier 1476 1492 positive labeled
T106 Reagent 1493 1498 cells
R37 Mod-Link Arg1:T106 Arg2:T105
T107 Action 1504 1513 assessing
E32 Action:T107 Measure-Type-Link:T108
T108 Measure-Type 1514 1530 purity and yield