protocol_156.ann

T1	Action 42 47	Place
E1	Action:T1 Acts-on:T23 Acts-on2:T24
T2	Action 155 160	added
E2	Action:T2 Acts-on:T30 Site:T28 Site2:T29
T3	Action 162 166	Spin
E3	Action:T3 Acts-on:T32
T4	Action 227 235	Pour off
E4	Action:T4 Acts-on:T35
T5	Action 249 252	Add
E5	Action:T5 Acts-on:T37 Site:T39
T6	Action 289 295	vortex
E6	Action:T6 Acts-on:T39
T7	Action 347 353	vortex
E7	Action:T7 Acts-on:T44
T8	Action 305 308	Add
E8	Action:T8 Acts-on:T43 Site:T44
T9	Action 371 375	spin
E9	Action:T9 
T10	Action 399 407	Transfer
E10	Action:T10 Acts-on:T53 Acts-on2:T50 Site:T54
T11	Action 460 470	Centrifuge
E11	Action:T11 Acts-on:T54
T12	Action 509 514	Empty
E12	Action:T12 Acts-on:T57
T13	Action 532 535	Add
E13	Action:T13 Acts-on:T59 Site:T60
T14	Action 569 573	Spin
E14	Action:T14 
T15	Action 612 615	Add
E15	Action:T15 Acts-on:T64 Site:T65
T16	Action 649 653	Spin
E16	Action:T16 
T17	Action 692 697	Place
E17	Action:T17 Acts-on:T68 Site:T69
T18	Action 739 743	spin
E18	Action:T18 
T19	Action 782 790	Transfer
E19	Action:T19 Acts-on:T73 Site:T75
T20	Action 845 848	Add
E20	Action:T20 Acts-on:T78 Site:T79
T21	Action 915 923	incubate
E21	Action:T21 
T22	Action 948 952	Spin
E22	Action:T22 
T23	Reagent 48 58	Buffer ATL
T24	Reagent 63 72	Buffer Al
T25	Temperature 76 79	56C
T26	Time 93 102	5 minutes
T27	Modifier 84 92	at least
R1	Mod-Link Arg1:T26 Arg2:T27	
R2	Setting Arg1:E1 Arg2:T25	
R3	Setting Arg1:E1 Arg2:T26	
T28	Reagent 130 133	AW1
T29	Reagent 138 141	AW2
T30	Reagent 147 154	ethanol
T31	Amount 167 173	>500ul
T32	Reagent 177 200	bacteria liquid culture
T33	Speed 204 210	10000g
T34	Time 215 225	10 minutes
R4	Measure Arg1:T32 Arg2:T31	
R5	Setting Arg1:E3 Arg2:T33	
R6	Setting Arg1:E3 Arg2:T34	
T35	Reagent 236 247	supernatant
T36	Amount 253 258	200ul
T37	Reagent 259 269	buffer ATL
T38	Time 300 303	15s
R7	Setting Arg1:E6 Arg2:T38	
T39	Reagent 278 284	sample
T40	Modifier 273 277	each
R8	Mod-Link Arg1:T39 Arg2:T40	
R9	Measure Arg1:T37 Arg2:T36	
T41	Amount 309 314	200ul
T42	Concentration 315 319	100%
T43	Reagent 320 327	ethanol
T44	Reagent 336 342	sample
T45	Modifier 331 335	each
T46	Time 358 361	15s
R10	Setting Arg1:E7 Arg2:T46	
R11	Measure Arg1:T43 Arg2:T41	
R12	Measure Arg1:T43 Arg2:T42	
R13	Mod-Link Arg1:T44 Arg2:T45	
T47	Modifier 363 370	Briefly
T48	Action 379 386	collect
E23	Action:T48 Acts-on:T49 Using:E9
T49	Reagent 391 397	liquid
R14	Mod-Link Arg1:E9 Arg2:T47	
T50	Reagent 415 421	sample
T51	Amount 387 390	all
R15	Measure Arg1:T49 Arg2:T51	
T52	Amount 408 411	all
R16	Measure Arg1:T50 Arg2:T52	
T53	Reagent 432 443	precipitate
T54	Location 447 458	spin column
T55	Speed 485 492	>20000g
T56	Time 498 507	2 minutes
R17	Setting Arg1:E11 Arg2:T55	
R18	Setting Arg1:E11 Arg2:T56	
T57	Location 515 530	collection tube
T58	Amount 536 541	500ul
T59	Reagent 542 552	buffer AW1
T60	Location 556 567	spin column
R19	Measure Arg1:T59 Arg2:T58	
T61	Speed 588 595	>20000g
T62	Time 601 610	2 minutes
R20	Setting Arg1:E14 Arg2:T61	
R21	Setting Arg1:E14 Arg2:T62	
T63	Amount 616 621	500ul
T64	Reagent 622 632	buffer AW2
T65	Location 636 647	spin column
R22	Measure Arg1:T64 Arg2:T63	
T66	Speed 668 675	>20000g
T67	Time 681 690	2 minutes
R23	Setting Arg1:E16 Arg2:T66	
R24	Setting Arg1:E16 Arg2:T67	
T68	Location 698 709	spin column
T69	Location 719 734	collection tube
T70	Modifier 715 718	new
T71	Speed 758 765	>20000g
T72	Time 771 780	3 minutes
R25	Mod-Link Arg1:T69 Arg2:T70	
R26	Setting Arg1:E18 Arg2:T71	
R27	Setting Arg1:E18 Arg2:T72	
T73	Location 791 802	spin column
T74	Amount 816 821	1.5ml
T75	Location 822 843	Eppendorf lobind tube
R28	Measure Arg1:T75 Arg2:T74	
T76	Modifier 808 815	labeled
R29	Mod-Link Arg1:T75 Arg2:T76	
T77	Amount 849 854	100ul
T78	Reagent 858 867	buffer TE
T79	Location 880 895	membrane/filter
T80	Location 899 910	spin column
T81	Temperature 927 936	room temp
R30	Setting Arg1:E21 Arg2:T81	
R31	Measure Arg1:T78 Arg2:T77	
R32	Meronym Arg1:T79 Arg2:T80	
T82	Time 942 947	1 min
T83	Speed 956 962	10000g
T84	Time 967 976	2 minutes
R33	Setting Arg1:E22 Arg2:T82	
R34	Setting Arg1:E22 Arg2:T83	
R35	Setting Arg1:E22 Arg2:T84