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protocol_307.ann
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T1 Action 46 53 Prepare
E1 Action:T1 Creates:T17
T2 Action 83 92 resuspend
E2 Action:T2 Acts-on:T18 Site:T19
T3 Action 174 178 Pass
E3 Action:T3 Acts-on:T20 Using:T22
T4 Action 213 223 centrifuge
E4 Action:T4 Acts-on:T20
T5 Action 249 256 discard
E5 Action:T5 Acts-on:T25
T6 Action 277 286 resuspend
E6 Action:T6 Acts-on:T26 Site:T27
T7 Action 324 330 Adjust
E7 Action:T7
T8 Action 375 382 Aliquot
E8 Action:T8 Acts-on:T33 Site:T32
T9 Action 419 425 Vortex
E9 Action:T9 Acts-on:T35
T10 Action 504 511 prepare
E10 Action:T10 Creates:T38 Using:E9
T11 Action 535 538 Add
E11 Action:T11 Acts-on:T40
T12 Action 582 585 Mix
E12 Action:T12
T13 Action 595 603 incubate
E13 Action:T13 Site:T44
T14 Action 627 635 Scale up
E14 Action:T14
T15 Action 698 701 add
E15 Action:T15 Acts-on:T48
T16 Action 921 930 Resuspend
E16 Action:T16 Acts-on:T51 Site:T54
T17 Reagent 56 78 single cell suspension
T18 Reagent 97 102 cells
T19 Reagent 108 139 ice cold cell separation buffer
T20 Reagent 183 188 cells
T21 Size 199 204 70 μm
T22 Device 205 211 filter
R1 Measure Arg1:T22 Arg2:T21
T23 Speed 225 232 300 x g
T24 Time 237 246 5 minutes
R2 Setting Arg1:E4 Arg2:T23
R3 Setting Arg1:E4 Arg2:T24
T25 Reagent 261 272 supernatant
T26 Reagent 291 296 cells
T27 Reagent 300 322 cell separation buffer
T28 Measure-Type 335 353 cell concentration
T29 Generic-Measure 357 373 1 x 108 cells/mL
R4 Of-Type Arg1:T29 Arg2:T28
R5 Setting Arg1:E7 Arg2:T29
T30 Amount 383 389 100 μL
T31 Modifier 409 412 new
T32 Location 413 417 tube
R6 Mod-Link Arg1:T32 Arg2:T31
T33 Reagent 395 400 cells
R7 Measure Arg1:T33 Arg2:T30
T34 Numerical 391 394 107
R8 Measure Arg1:T33 Arg2:T34
T35 Reagent 430 459 antibody-conjugated Nanobeads
T36 Speed 478 487 max speed
T37 Generic-Measure 489 498 5 touches
T38 Reagent 516 525 dilutions
R9 Setting Arg1:E9 Arg2:T36
R10 Setting Arg1:E9 Arg2:T37
T39 Amount 539 544 10 μL
T40 Reagent 571 580 Nanobeads
R11 Measure Arg1:T40 Arg2:T39
T41 Modifier 548 559 pre-diluted
T42 Modifier 560 570 conjugated
R12 Mod-Link Arg1:T40 Arg2:T42
R13 Mod-Link Arg1:T40 Arg2:T41
T43 Modifier 586 590 well
T44 Location 607 610 ice
T45 Time 615 625 15 minutes
R14 Mod-Link Arg1:E12 Arg2:T43
R15 Setting Arg1:E13 Arg2:T45
T46 Amount 702 708 100 μL
T47 Modifier 712 723 pre-diluted
T48 Reagent 724 733 Nanobeads
T49 Generic-Measure 738 745 1 x 108
T50 Reagent 746 751 cells
R16 Measure Arg1:T50 Arg2:T49
R17 Mod-Link Arg1:T48 Arg2:T47
R18 Measure Arg1:T48 Arg2:T46
T51 Reagent 935 940 cells
T52 Modifier 944 955 appropriate
T53 Amount 956 962 amount
T54 Reagent 966 972 buffer
R19 Measure Arg1:T54 Arg2:T53
R20 Mod-Link Arg1:T53 Arg2:T52