# Array dev- part 2 of the filter Sand Hybe process # ESP DELTA unit with new valve manifold # rev 015a; July 20 2002 # Edited from GOM macro # Starts with ~1.5 ml lysate from sandhybe1 macro # Syringe begins @ pos 5 mL # valve config 001 # no delays between steps now reduce 10 minutes # clear filter between washes with syringe pull # increased push pulls signal/conj fills # reduced con wash preeure strokes # incl. Lysis syringe rinse to kill conj B/4 add sub # 15 vs 15a: changed length of time for heating # 15 vs 15a: changed final sub push from 3.5 to 3.25 to prevent fart # 15 vs 15a: sample hybe temp at 30C # 15b - use "Signal2" as different wash buffer for O/N run PROGRAM: SH2 HEAT 39.75 63 # puck T=~30C; check T delay=process t DELAYMIN 1 # pre-warm array puck SYRINGE PUSH 1.25 Puck VALVEPORT Puck HOME SYRINGEJOG PUSH 2.85 SYRINGEJOG PULL 2.85 SYRINGE PUSH ALL ToxicWaste DELAYMIN 2 SYRINGE PULL 0.5 Air # try push/pull 1.5 to prevent foaming SYRINGE PUSH 0.5 Air DELAYMIN 2 # 2nd lysate push SYRINGE PULL 0.5 Air SYRINGE PUSH 0.5 Air DELAYMIN 2 # 3rd lysate push SYRINGE PULL 0.5 Air SYRINGE PUSH 0.5 Air DELAYMIN 2 # 4th lysate push SYRINGE PULL 0.5 Air SYRINGE PUSH 0.5 Air DELAYMIN 2 # 5th lysate push; hybe t=10 min SYRINGE PULL 3.5 Air # clear lysate SYRINGE PUSH ALL ToxicWaste SYRINGE PULL 1.5 M3Bypass SYRINGE PUSH ALL ToxicWaste # burp M1 loop;ready for wash SYRINGE PULL 2.5 Signal2 # SIGNAL 2 = WASH WITH 0.5% TWEEN VALVEPORT Signal2 HOME SYRINGEJOG PULL 2 SYRINGE PUSH 1.5 Puck # prime connection to filter VALVEPORT Puck HOME # isolate filter SYRINGEJOG PUSH 1.85 # fill filter holder DELAYSEC 55 # lysate wash 1 of 1 SYRINGE PUSH ALL Puck SYRINGE PULL 3.5 Air # air out puck SYRINGE PUSH ALL ToxicWaste # puck cleared SYRINGE PULL 1.5 M3Bypass SYRINGE PUSH ALL ToxicWaste # burp M1 loop;ready for sig SYRINGE PULL 2 Signal1 VALVEPORT Signal1 HOME SYRINGEJOG PULL 2 # clear mnfld SYRINGE PUSH 1.5 Puck VALVEPORT Puck HOME SYRINGEJOG PUSH 1.85 SYRINGEJOG PULL 1.85 VALVEPORT Puck SET VALVEPORT Puck HOME # puck filled with sig probe SYRINGE PUSH ALL ToxicWaste SYRINGE PULL 0.75 Air SYRINGE PUSH 0.75 Air # array bkflsh DELAYMIN 2 # equilibrate array with sig SYRINGE PULL 3.5 Air # air out puck SYRINGE PUSH ALL ToxicWaste # puck cleared SYRINGE PULL 1.5 M3Bypass SYRINGE PUSH ALL ToxicWaste # burp M1 loop;ready for sig SYRINGE PULL 2 Signal1 VALVEPORT Signal1 HOME SYRINGEJOG PULL 2 # clear mnfld SYRINGE PUSH 1.5 Puck VALVEPORT Puck HOME SYRINGEJOG PUSH 1.85 SYRINGEJOG PULL 1.85 VALVEPORT Puck SET VALVEPORT Puck HOME # puck filled with sig probe SYRINGE PUSH ALL ToxicWaste SYRINGE PULL 0.75 Air SYRINGE PUSH 0.75 Air # array bkflsh DELAYMIN 2 SYRINGE PULL 0.75 Air SYRINGE PUSH 0.75 Air # check push/pull vol DELAYMIN 2 SYRINGE PULL 0.75 Air SYRINGE PUSH 0.75 Air DELAYMIN 2 SYRINGE PULL 0.75 Air SYRINGE PUSH 0.75 Air DELAYMIN 2 # sig probe hybe complete t=10 min SYRINGE PULL 3.5 Air # air out puck SYRINGE PUSH ALL ToxicWaste # puck cleared SYRINGE PULL 1.5 M3Bypass SYRINGE PUSH ALL ToxicWaste # burp M1 loop;ready for wash1 SYRINGE PULL 2.5 Signal2 VALVEPORT Signal2 HOME SYRINGEJOG PULL 2 SYRINGE PUSH 1.5 Puck # prime connection to filter VALVEPORT Puck HOME # isolate filter SYRINGEJOG PUSH 1.75 # fill filter holder DELAYSEC 55 # sig probe wash 1 of 2 SYRINGE PUSH ALL Puck SYRINGE PULL 2.5 Signal2 VALVEPORT Signal2 HOME SYRINGEJOG PULL 2 SYRINGE PUSH 1.5 Puck # prime connection to filter VALVEPORT Puck HOME # isolate filter SYRINGEJOG PUSH 1.75 # fill filter holder DELAYSEC 55 # sig probe wash 2 of 2 SYRINGE PUSH ALL Puck SYRINGE PULL 3.5 Air # air out puck SYRINGE PUSH ALL ToxicWaste # puck cleared SYRINGE PULL 1.5 M3Bypass SYRINGE PUSH ALL ToxicWaste # burp M1 loop;ready for con SYRINGE PULL 2 Conjugate VALVEPORT Conjugate HOME SYRINGEJOG PULL 2 # clear mnfld SYRINGE PUSH 1.5 Puck VALVEPORT Puck HOME SYRINGEJOG PUSH 1.85 SYRINGEJOG PULL 1.85 VALVEPORT Puck SET VALVEPORT Puck HOME # puck filled with con SYRINGE PUSH ALL ToxicWaste SYRINGE PULL 0.75 Air SYRINGE PUSH 0.75 Air # array bkflsh DELAYMIN 2 # equilibrate array with con SYRINGE PULL 3.5 Air # air out puck SYRINGE PUSH ALL ToxicWaste # puck cleared SYRINGE PULL 1.5 M3Bypass SYRINGE PUSH ALL ToxicWaste # burp M1 loop;ready for con SYRINGE PULL 2 Conjugate VALVEPORT Conjugate HOME SYRINGEJOG PULL 2 # clear mnfld SYRINGE PUSH 1.5 Puck VALVEPORT Puck HOME SYRINGEJOG PUSH 1.85 SYRINGEJOG PULL 1.85 VALVEPORT Puck SET VALVEPORT Puck HOME # puck filled with con SYRINGE PUSH ALL ToxicWaste DELAYMIN 2 # isolate puck with con SYRINGE PULL 0.75 Air SYRINGE PUSH 0.75 Air # array bkflsh DELAYMIN 2 SYRINGE PULL 0.75 Air SYRINGE PUSH 0.75 Air DELAYMIN 2 # con complete t=8min SYRINGE PULL 3.5 Air # air out puck SYRINGE PUSH ALL ToxicWaste # puck cleared SYRINGE PULL 1.5 M3Bypass SYRINGE PUSH ALL ToxicWaste # burp M1 loop;ready for con wash1 SYRINGE PULL 2 Signal2 # Vector Buffer E VALVEPORT Signal2 HOME SYRINGEJOG PULL 2 # clear mnfld SYRINGE PUSH 1.5 Puck # prime connection to puck VALVEPORT Puck HOME SYRINGEJOG PUSH 1.7 SYRINGEJOG PULL 1.7 # relieve pressure VALVEPORT Puck SET # prime lower loop VALVEPORT Puck HOME # puck filled with wash fluid SYRINGE PUSH ALL ToxicWaste DELAYSEC 10 SYRINGE PULL 0.75 Air SYRINGE PUSH 0.75 Air # array bkflsh DELAYSEC 10 # end con wash 1 SYRINGE PULL 4 Air # air out puck SYRINGE PUSH ALL ToxicWaste # puck cleared SYRINGE PULL 1.5 M3Bypass SYRINGE PUSH ALL ToxicWaste # burp M1 loop;ready for con wash 2 SYRINGE PULL 2 Signal2 # Vector Buffer E VALVEPORT Signal2 HOME SYRINGEJOG PULL 2 # clear mnfld SYRINGE PUSH 1.5 Puck # prime connection to puck VALVEPORT Puck HOME SYRINGEJOG PUSH 1.7 SYRINGEJOG PULL 1.7 # relieve pressure VALVEPORT Puck SET # prime lower loop VALVEPORT Puck HOME # puck filled with wash fluid SYRINGE PUSH ALL ToxicWaste DELAYSEC 10 SYRINGE PULL 0.75 Air SYRINGE PUSH 0.75 Air # array bkflsh DELAYSEC 10 # end con wash 2 SYRINGE PULL 4 Air # air out puck SYRINGE PUSH ALL ToxicWaste # puck cleared SYRINGE PULL 1.5 M3Bypass SYRINGE PUSH ALL ToxicWaste # burp M1 loop;ready for con wash 3 SYRINGE PULL 2 Signal2 # Vector Buffer E VALVEPORT Signal2 HOME SYRINGEJOG PULL 2 # clear mnfld SYRINGE PUSH 1.5 Puck # prime connection to puck VALVEPORT Puck HOME SYRINGEJOG PUSH 1.7 SYRINGEJOG PULL 1.7 # relieve pressure VALVEPORT Puck SET # prime lower loop VALVEPORT Puck HOME # puck filled with wash fluid SYRINGE PUSH ALL ToxicWaste DELAYSEC 10 SYRINGE PULL 0.75 Air SYRINGE PUSH 0.75 Air # array bkflsh DELAYSEC 10 # end con wash 3 SYRINGE PULL 4 Air # air out puck SYRINGE PUSH ALL ToxicWaste # puck cleared SYRINGE PULL 1.5 M3Bypass SYRINGE PUSH ALL ToxicWaste # burp M1 loop;ready for Kill step SYRINGE PULL 2 Lysis # KILL CON IN SYRINGE DELAYSEC 10 VALVEPORT Lysis HOME SYRINGEJOG PULL 3 DELAYSEC 10 SYRINGE PUSH ALL ToxicWaste SYRINGE PULL 5 FlushM2 VALVEPORT FlushM2 HOME SYRINGEJOG PULL 4 # clears out manifold SYRINGE PUSH ALL ToxicWaste # ends lysis poison SYRINGE PULL 1.5 M3Bypass SYRINGE PUSH ALL ToxicWaste # burp M1 loop CAMERAPOWER ON # START CAMERA COOLING SYRINGE PULL 4 FlushM2 VALVEPORT FlushM2 HOME SYRINGEJOG PULL 4 SYRINGE PUSH ALL ToxicWaste SYRINGE PULL 1.5 M2Bypass # pull in air to blow remaining fluid SYRINGE PUSH ALL ToxicWaste # flushed; ready for sub SYRINGE PULL 1.0 Sub1 SYRINGE PULL 1.0 Sub2 VALVEPORT Sub2 HOME SYRINGEJOG PULL 4 # mix sub 1&2 SYRINGE PUSH 1.5 Puck VALVEPORT Puck HOME SYRINGEJOG PUSH 3.25 # CK VOL TO MAKE SURE ALL IS IN PUCK! DELAYSEC 10 SYRINGE PUSH ALL Puck SYRINGE PULL 5 Air # air out puck for imaging ACQUIREIMAGE LIGHT, 2X2, 60000, "1 min exp" ACQUIREIMAGE LIGHT, 2X2, 240000, "4 min exp" CAMERAPOWER OFF #SHUT CAMERA DOWN SYRINGE PUSH ALL ToxicWaste END