Blood/Cell Culture Extraction Handbook

Transcription

1 ABIOpure T M Genmic DNA Bld/Cell Culture Extractin Handbk Cat N: M501DP FOR RESEARCH USE ONLY

2 Table f Cntent Cntent Page Kit Cmpnents 4 Cautin 4 Stability & Strage 4 General Descriptin 5 Limitatins 5 Quality Cntrl 5 Technical Supprt 6 Warning & Precautins 6 Features 7 Tested data 7 Samples 8 Quick Prtcl 9 Reagents Preparatin 10 Materials required but nt prvided 10 Bld Prtcl 11 Befre yu start! 11 Prcedure 12 A. RBC Lysis Fresh Bld 12 A. RBC Lysis Frzen Bld 13 B. Cell Lysis 14 May

3 Cntent Page C. DNA Binding 14 D. Washing 15 E. Elutin 15 Buffy Cat Prtcl 16 Befre yu start! 16 Prcedure 17 A. RBC Lysis 17 B. Cell Lysis 18 C. DNA Binding 18 D. Washing 19 E. Elutin 19 Cultured Cells and Nn-human bld Prtcl 20 Befre yu start! 20 Prcedure 21 A. Sample preparatin and Cell Harvesting 21 Cultured Animal Cells 21 Gram - Negative Bacteria 21 Gram - Psitive Bacteria 22 Fungal Cells 22 Fresh Bld (ther than human) 23 B. Cell Lysis 23 May

4 Cntent Page C. DNA Binding 24 D. Washing 24 E. Elutin 25 Trubleshting 26 Ordering Infrmatin 27 May

5 Kit Cmpnents Cmpnent M501DP04 M501DP100 M501DP300 N. f Preps RBC Lysis Buffer 6 ml 135 ml 405 ml GT Buffer 1.5 ml 30 ml 75 ml GB Buffer 2 ml 40 ml 100 ml W1 Buffer 2 ml 45 ml 130 ml Wash Buffer * (add Ethanl) 1 ml (4 ml) 25 ml (100 ml) 50 ml (200 ml) Elutin Buffer 1 ml 30 ml 75 ml GD Clumn 4 pcs 100 pcs 300 pcs 2 ml Cllectin Tube 8 pcs 200 pcs 600 pcs * Add abslute ethanl t the Wash Buffer prir t initial use (see the bttle label fr vlume). Cautin GB Buffer cntains guanidine hydrchlride that is a harmful irritant. During peratin always wear a lab cat, dispsable glves and prtective gggles. Stability and strage ABIOpure TM Extractin kits are stable fr 2 years when stred at rm temperature (15 25 C) in dry cnditins. May

6 General Descriptin ABIOpure TM Genmic DNA Bld/Cell Culture Extractin Kit prvides an efficient methd fr purifying ttal DNA (genmic, mitchndrial and viral DNA) frm whle bld, frzen bld, buffy cat, cultured cells and fungi. Chatrpic salt is used t lyse cells and degrade prtein. DNA is bund t the spin clumn, cntaminants are remved using a Wash Buffer and the purified genmic DNA is eluted by a lw salt Elutin Buffer r TE. The entire prcedure can be cmpleted in less than an hur withut phenl/chlrfrm extractin r alchl precipitatin, with an average yield f 6μg frm 200μl f whle human bld and up t 50µg f DNA frm 200µl f buffy cat. Limitatin The ABIOpure TM Genmic DNA Extractin kit is intended fr mlecular bilgy applicatins. This prduct is nt intended fr the diagnsis r treatment f diseases. Quality Cntrl The Alliance Bi Quality Cntrl department perfrms stringent quality cntrl prcedures n every lt f ABIOpure TM Extractin kits prduced and keeps cmplete recrds f ur prductin prcess fr lt tracking and after sale trubleshting and supprt. May

7 Technical Supprt The Alliance Bi Technical Supprt team has decades f experience in Life Sciences and rich scientific backgrund in mlecular applicatins. If yu need assistance, have any questin r suggestin r if yu experience any difficulties using ABIOpure TM Extractin kits, please feel free t cntact ur technical supprt team at supprt@alliancebi.cm. The Alliance Bi Technical Supprt department and R&D lab wrk tgether t enhance the prducts perfrmance t match ur custmers need and satisfactin. Warning and Precautins When wrking with chemicals, always wear a suitable lab cat, dispsable glves, and prtective gggles. Handle and dispse f all bilgical samples as if they were able t transmit infective agents. Avid direct cntact with the bilgical samples. Avid prducing spills r aersl. Any material cming in cntact with the bilgical samples must be treated fr at least 30 minutes with 3% sdium hypchlrite r autclaved fr ne hur at 121 C befre dispsal. Never pipette slutins by muth! Handle and dispse ff all reagents and all materials used t carry ut the assay as if they were able t transmit infective agents. Avid direct cntact with the reagents. Avid prducing spills r aersl. Waste must be handled and dispsed f accrding t adequate safety measures. Dispsable cmbustible material must be incinerated. Samples and reagents must be handled under Laminar flw hd. Always wrk in cmpliance with GLP (Gd Labratry Practice) standards. May

8 Features Frmat: Spin Clumn Yield: up t 50µg Operatin time: frm 35 60min (accrding t prtcl fllwed) Elutin Vlume: µl Applicatins: PCR, Sequencing and varius ther enzymatic reactins Tested Data 300µl f whle bld was used fr this test. The final prduct was eluted in 200µl f Elutin Buffer and 15µl f the final sample was laded int each lane (M: 1Kb DNA Ladder). M ABIOpure TM Genmic DNA Bld/Cell Culture Extractin Kit was tested fr perfrmance against cmpetitrs and shwed equal t superir perfrmance against leading cmpetitrs. May

9 Samples: Up t 300µl f Whle Bld Up t 200µl f Frzen Whle Bld Up t 200µl f Buffy Cat (up t 5 x 10 6 ) Cultured animal cells (up t 1 X 10 7 ) Bacteria (up t 1 X 10 9 ) Fungi (up t 5 X 10 7 ) Fr DNA purificatin frm mammalian bld ther than human (nucleated and nn-nucleated erythrcytes), ABIOpure TM Genmic DNA Bld/Cell Culture Extractin Kit Cultured Cell prtcl is recmmended. Fr purificatin f DNA frm tissue, paraffin-embedded tissue, buccal swab and amnitic fluid, ABIOpure TM Genmic DNA Tissue Extractin Kit is recmmended. Fr purificatin f DNA frm fresh plant tissue and dry plant tissue, ABIOpure TM Genmic DNA Plant Extractin Kit is recmmended. Fr purificatin f viral DNA/RNA frm plasma, serum, bdy fluids and viral infected cell cultures, ABIOpure TM Viral Extractin Kit and ABIOpure TM Viral Plus Extractin Kit are recmmended. May

10 Quick Prcedure RBC Lysis (add RBC lysis t Bld samples) Cell Lysis (Add GB Buffer) DNA Binding (Transfer t GD clumn) Washing (Wash with W1 and then with Wash Buffer) Elutin (Elute with Elutin Buffer r TE) May

11 Reagents Preparatin Reagents included are ready t use except fr the Wash Buffer. Add abslute ethanl (see the bttle label fr vlume) t the Wash Buffer prir t initial use and tick the mark n the bttle that the ethanl has been added. Materials required but nt prvided Laminar flw hd Dispsable glves Ethanl (96 100%) 1.5 r 2.0ml micrcentrifuge tubes Pipette set (10µl, 100µl and 1000µl) Pipette tips with aersl barriers Micrcentrifuge Vrtex Water Bath r heating blck (up t 70 C) PBS (Phsphate-buffered saline) fr certain samples Fr Bacteria Prtcl: lyszyme buffer (20mg/ml lyszyme; 20mM Tris-HCl; 2mM EDTA; 1% Tritn X-100; ph 8.0) Fr Fungus Prrcl: lyticase r zymlase, srbitl buffer (1.2 M srbitl; 10mM CaCl2; 0.1M Tris-HCl ph 7.5; 35mM mercaptethanl) * Prteinase K * RNase A (10mg/ml) * Optinal May

12 Bld Prtcl This prtcl is intended fr the extractin f ttal genmic, mitchndrial and/r viral DNA frm fresh and frzen Whle Bld samples. Befre yu start! RBC Lysis Buffer is prvided t remve nn-nucleated red bld cells and reduce hemglbin cntaminatin. When the bld sample is less than 5μl r the sample cnsists f nucleated bld cells, we recmmend using the Cultured Cell Prtcl t purify genmic DNA. 200μl f whle human bld yield f 6μg f DNA. If higher yield is required, Buffy cat preparatin is recmmended t reach a yield f up t 50µg f DNA. Buffy cat prcedure is recmmended fr higher perfrmance. Centrifugatin steps t be carried ut at rm temperature (15 25 C). Let yur samples stand fr few minutes t reach rm temperature. Adjust yur water bath r heating blck at 60 C. Ensure that the Wash Buffer has been suspended with abslute ethanl accrding t the vlume indicated n the bttle. Preheat the required Elutin Buffer (50-200µl per sample) in a 70 C water bath fr the elutin step. Fr all type f samples; standard elutin vlume is 100µl. If less sample vlume is used, reduce the elutin vlume t increase DNA cncentratin. Fr higher yield, repeat the Elutin step with further 100µl Elutin Buffer t increase recvery. May

13 Bld Prtcl - Cntinue Prcedure Cllect bld samples in EDTA-Na 2 treated cllectin tubes (r ther anticagulant mixture that des nt affect yur later applicatin). Start the extractin prcedure within 4 hurs after cllectin r stre at 4 C fr up t 3 days. Fr lng-term strage, it is recmmended t stre the bld samples at -70 C. A. RBC Lysis Fresh Bld a1. Cllect fresh bld in EDTA-Na 2 treated cllectin tubes (r ther anticagulant mixture that des nt affect yur later applicatin). a2. Transfer up t 300µl f fresh bld t a 1.5 ml micrcentrifuge tube. If the bld sample is mre than 300µl (up t 1 ml), add the sample t a sterile 15ml centrifuge tube. If the sample is less than 300µl, add the apprpriate vlume f PBS. a3. Add 3x the sample vlume f RBC Lysis Buffer and mix by inversin. D nt vrtex. a4. Incubate the tube fr 10 minutes at rm temperature. a5. Centrifuge fr 5 minutes at 3000 x g and remve the supernatant cmpletely. a6. Add 100µl f RBC Lysis Buffer t re-suspend the cell pellet. a7. Prceed with Cell Lysis (step n. b1) in the fllwing table. May

14 Bld Prtcl - Cntinue Frzen Bld a1. Frzen bld shuld be thawed in a 37 C water bath with mild agitatin befre beginning the prcedure. a2. Bring yur samples t rm temperature. a3. Add up t 200µl f bld t a 1.5ml micrcentrifuge tube. If the sample vlume is less than 200 µl add the apprpriate vlume f PBS. a4. Add 30µl f Prteinase K (10mg/ml) t the 1.5ml micrcentrifuge tube and mix briefly. Incubate the mixture at 60 C fr 15 minutes. During incubatin, invert tube every 3 minutes. a5. Prceed with Cell Lysis (step n. b1) in the fllwing table. May

15 C. DNA Binding B. Cell Lysis Bld Prtcl - Cntinue b1. After RBC Lysis step, add 200µl f GB Buffer t the 1.5ml micrcentrifuge tube cntaining the sample (described abve) and mix by vrtex. b2. Incubate at 60 C fr 10 min r until the sample lysate is clear. (Invert the tube every 3 min) ** Fr Frzen bld samples, it s preferable t incubate at 70 C fr 15 minutes. (Invert the tube every 3 min) Optinal Step: RNA Degradatin If RNA-free genmic DNA is required, perfrm this ptinal step. Add 5µl f RNase A (10mg/ml) t the sample lysate and mix by vrtex. Incubate at rm temperature fr 5 minutes. c1. Add 200µl f abslute ethanl t the sample lysate and vrtex immediately fr 10 secnds. If precipitate appears, break it up by pipetting. c2. Place a GD Clumn in a 2ml Cllectin Tube and transfer all f the mixture (including any precipitate) t the GD Clumn. c3. Centrifuge at full speed fr 5 minutes. c4. Discard the 2ml Cllectin Tube cntaining the flw-thrugh and place the GD Clumn in a new 2ml Cllectin Tube. May

16 E. Elutin D. Washing Bld Prtcl - Cntinue d1. Add 400µl f W1 Buffer t the GD Clumn. d2. Centrifuge at full speed fr 30 secnds. d3. Discard the flw-thrugh and place the GD Clumn back in the 2ml Cllectin Tube. d4. Add 600µl f Wash Buffer (ethanl added) t the GD Clumn. d5. Centrifuge at full speed fr 30 secnds. d6. Discard the flw-thrugh and place the GD Clumn back in the 2ml Cllectin Tube. Recmmended: Centrifuge again fr 3 minutes at full speed t dry the clumn matrix. e1. Transfer the dried GD Clumn t a clean 1.5ml micrfuge tube. e2. Add (50 200µl) f preheated Elutin Buffer r TE t the center f the clumn matrix. e3. Centrifuge at full speed fr 1 minute t elute the purified DNA. Fr higher yield, let the GD clumns stand fr 3 5 minutes befre centrifugatin. Prceed with yur desired applicatin r stre at -20 C. May

17 Buffy Cat Prtcl This prcedure is intended fr the extractin f ttal genmic, mitchndrial and/r viral DNA frm Buffy Cat samples. Buffy cat is a leukcyte-enriched fractin f anticagulated bld sample after density gradient centrifugatin. Prepare buffy cat by centrifuging whle bld sample at 2500 x g fr 10 minutes at rm temperature. After centrifugatin, cllect the buffy layer (intermediate layer) cntaining cncentrated leukcytes withut interfering with the upper r lwer layers. Using buffy-cat frm whle bld yields higher DNA cncentratin than an equivalent vlume f whle bld. Befre yu start! Centrifugatin steps t be carried ut at rm temperature (15 25 C). Let yur samples stand fr few minutes t reach rm temperature. Adjust yur water bath r heating blck at 60 C. Ensure that the Wash Buffer has been suspended with abslute ethanl accrding t the vlume indicated n the bttle. Preheat the required Elutin Buffer (50-200µl per sample) in a 70 C water bath fr the elutin step. Standard elutin vlume is 100µl. If less sample vlume is used, reduce the elutin vlume t increase DNA cncentratin. Fr higher yield, repeat the Elutin step with further 100µl Elutin Buffer t increase recvery. May

18 A. RBC Lysis Buffy cat Prtcl Cntinue Prcedure Fllw standard Buffy cat preparatin prcedures then prceed with fllwing the steps: a1. Transfer up t 200µl f buffy cat (n mre than 5 x 10 6 f cells) t a 1.5 ml micrcentrifuge tube. a2. Add 3x the sample vlume f RBC Lysis Buffer t the tube and mix by inversin. a3. Incubate the tube fr 10 minutes at rm temperature. During incubatin, invert the tube every 3 minutes. a4. Centrifuge at full speed fr 1 minute and discard the supernatant cmpletely. a5. Add 500µl f RBC Lysis Buffer t re-suspend the white pellet. Mix by pipetting up and dwn. a6. Centrifuge at full speed fr 1 minute and discard the supernatant cmpletely. a7. Add 200µl f RBC Lysis Buffer t the tube and re-suspend the white pellet cmpletely by pipetting up and dwn. Mix the tube by vrtex nly if the pellet is nt cmpletely resuspended. May

19 C. DNA Binding B. Cell Lysis Buffy cat Prtcl Cntinue b1. Add 250µl f GB Buffer t the 1.5ml micrcentrifuge tube and mix by vrtex. b2. Incubate at 60 C fr 30 minutes r until the sample lysate is clear. (Invert the tube every 3 min). Optinal Step: RNA Degradatin If RNA-free genmic DNA is required, perfrm this ptinal step. Add 5µl f RNase A (10mg/ml) t the sample lysate and mix by vrtex. Incubate at rm temperature fr 5 minutes. c1. Add 250µl f abslute ethanl t the sample lysate and vrtex immediately fr 10 secnds. If precipitate appears, break it up by pipetting. c2. Place a GD Clumn in a 2ml Cllectin Tube and transfer all f the mixture (including any precipitate) t the GD Clumn. c3. Centrifuge at full speed fr 5 minutes. c4. Discard the 2ml Cllectin Tube cntaining the flwthrugh and place the GD Clumn in a new 2ml Cllectin Tube. May

20 E. Elutin D. Washing Buffy cat Prtcl Cntinue d1. Add 400µl f W1 Buffer t the GD Clumn. d2. Centrifuge at full speed fr 30 secnds. d3. Discard the flw-thrugh and place the GD Clumn back in the 2ml Cllectin Tube. d4. Add 600µl f Wash Buffer t the GD Clumn and centrifuge at full speed fr 30 secnds. d5. Centrifuge at full speed fr 30 secnds. d6. Discard the flw-thrugh and place the GD Clumn back in the 2ml Cllectin Tube. Recmmended: Centrifuge again at full speed fr 3 minutes t dry the clumn matrix. e1. Transfer the dried GD Clumn t a clean 1.5ml micrfuge tube. e2. Add (50-200µl) f preheated Elutin Buffer r TE t the center f the clumn matrix. e3. Centrifuge at full speed fr 1 minute t elute the purified DNA. Fr higher yield, let the GD clumns stand fr 3 5 minutes befre centrifugatin. Prceed with yur desired applicatin r stre at -20 C. May

21 Cultured Cells and Nn-human Bld Prtcl This prtcl is intended fr the extractin f ttal genmic, mitchndrial and/r viral DNA frm animal cultured cells, gram-psitive bacteria, gram-negative bacteria, fungal cells, mammalian fresh bld ther than human (nn-nucleated erythrcytes) and mammalian fresh bld with nucleated erythrcytes (e.g. fish r bird). Befre yu start! Centrifugatin steps t be carried ut at rm temperature (15 25 C). Let yur samples stand fr few minutes t reach rm temperature. Warm up yur water bath r heating blck accrding t the starting temperature f the chsen prcedure. Ensure that the Wash Buffer has been suspended with abslute ethanl accrding t the vlume indicated n the bttle. Preheat the required Elutin Buffer (30-200µl per sample) in a 70 C water bath fr the elutin step. Fr Gram-psitive bacteria, lyszyme slutin is needed. 20mg/ml lyszyme; 20mM Tris-HCl; 2mM EDTA; 1% Tritn X-100; ph 8.0. Prepare fresh lyszyme slutin immediately prir t use. Fr Fungal cells, lyticase r zymlase, and srbitl buffer (1.2M srbitl; 10mM CaCl2; 0.1M Tris-HCl ph 7.5; 35mM mercaptethanl) shuld be ready fr sample preparatin. May

22 Cultured Cells and Nn-human Bld Prtcl Cntinue Prcedure A. Sample Preparatin and Cell Harvesting Cultured Animal Cells If using adherent cells, trypsinize the cells befre harvesting. a1. Transfer cells (up t 1 x 10 7 ) t a 1.5ml micrcentrifuge tube and harvest with centrifugatin fr 20 secnds at 6000 x g. a2. Discard the supernatant and re-suspend the cells with 150µl f RBC Lysis Buffer. a3. Prceed with Cell Lysis (step n. b1) in the fllwing table. Gram-negative bacteria a1. Transfer cultured bacterial cells (up t 1x10 9 ) t a 1.5ml micrcentrifuge tube. a2. Centrifuge fr 1 minute at full speed and discard the supernatant a3. Add 200µl f GT Buffer t the tube and re-suspend the cell pellet by vrtex r pipetting. a4. Incubate at rm temperature fr 5 minutes. a5. Prceed with Cell Lysis (step n. b1) in the fllwing table. Cultured Cells and Nn-human Bld Prtcl Cntinue May

23 Cultured Cells and Nn-human Bld Prtcl Cntinue Gram-psitive bacteria a1. Transfer cultured bacterial cells (up t 1x10 9 ) t a 1.5ml micrcentrifuge tube. a2. Centrifuge fr 1 minute at full speed and discard the supernatant. a3. Add 200μl f lyszyme buffer t the tube and re-suspend the cell pellet by vrtex r pipetting. a4. Incubate at rm temperature fr 10 minutes. During incubatin, invert the tube every 2-3 minutes. a5. Prceed with Cell Lysis (step n. b1) in the fllwing table. Fungal Cells a1. Harvest fungus cells (up t 5x10 7 ) by centrifugatin fr 10 minutes at 5000 x g. a2. Discard the supernatant and resuspend the pellet in 600µl f srbitl buffer. a3. Add 200U f lyticase r zymlase. a4. Incubate at 30 C fr 30 minutes. a5. Centrifuge the mixture fr 10 minutes at 2000 x g t harvest the spherplast. a6. Remve the supernatant and add 200μl f GT Buffer t the tube and re-suspend the cell pellet by vrtex r pipetting. a7. Incubate at rm temperature fr 5 minutes. a8. Prceed with Cell Lysis (step n. b1) in the fllwing table. May

24 B. Cell Lysis Cultured Cells and Nn-human Bld Prtcl Cntinue Fresh Bld (frm ther than human) Fr mammalian bld (nn-nucleated), the sample vlume can be up t 50µl. Fr nucleated erythrcytes (eg. bird r fish), the sample vlume can be up t 10µl. a1. Add 150µl f GT Buffer t a 1.5 ml micrcentrifuge tube alng with the bld sample and mix by shaking vigrusly r by vrtex. a2. Prceed with Cell Lysis (step n. b1) in the fllwing table. b1. Add 200µl f GB Buffer t the 1.5ml micrcentrifuge tube and mix by vrtex fr 5 secnds. b2. Incubate at 70 C fr 10 minutes r until the sample lysate is clear. During incubatin, invert the tube every 3 minutes. Optinal Step: RNA Degradatin If RNA-free genmic DNA is required, perfrm this ptinal step. After 70 C incubatin, add 5µl f RNase A (10mg/ml) t the sample lysate and mix by vrtex. Incubate at rm temperature fr 5 minutes. May

25 D. Washing C. DNA Binding Cultured Cells and Nn-human Bld Prtcl Cntinue c1. Add 200µl f abslute ethanl t the sample lysate and vrtex immediately fr 10 secnds. If precipitate appears, break it up by pipetting. c2 Place a GD Clumn in a 2ml Cllectin Tube and transfer all f the mixture (including any precipitate) t the GD Clumn. c3. Centrifuge at full speed fr 2 minutes. c4. Discard 2ml Cllectin Tube cntaining the flw-thrugh and place the GD Clumn in a new 2ml Cllectin tube. d1. Add 400µl f W1 Buffer t the GD Clum. d2. Centrifuge at full speed fr 30 secnds. d3. Discard the flw-thrugh and place the GD Clumn back in the 2ml Cllectin Tube. d4. Add 600µl f Wash Buffer t the GD Clumn. d5. Centrifuge at full speed fr 30 secnds. d6. Discard the flw-thrugh and place the GD Clumn back in the 2ml Cllectin Tube. Recmmended: Centrifuge again fr 3 minutes at full speed t dry the clumn matrix. May

26 E. Elutin Cultured Cells and Nn-human Bld Prtcl Cntinue e1. Standard elutin vlume is 100 µl. If less sample vlume is used, reduce the elutin vlume (30 50 µl) t increase DNA cncentratin. Fr higher yield, repeat the Elutin step t increase recvery and the ttal elutin vlume t apprximately 200 µl. Transfer the dried GD Clumn t a clean 1.5 ml micrfuge tube. e2. Add 100µl f preheated Elutin Buffer r TE t the center f the clumn matrix. e3. Centrifuge at full speed fr 1 minute t elute the purified DNA. Fr higher yield, let the GD clumns stand fr 3 5 minutes befre centrifugatin. Prceed with yur desired applicatin r stre at -20 C. May

27 Trubleshting Prblem Clgged Clumn Lw Yield Pssible Reasns/ Slutin T much sample was used. Reduce sample vlume r separate int multiple tubes. Precipitate was frmed at DNA Binding Step Reduce the sample material. Prir t lading the clumn, break up precipitate in ethanl-added lysate. Incrrect DNA Elutin Step Ensure that the Elutin Buffer r TE is added t the center f the GD Clumn matrix and is absrbed cmpletely. Incmplete DNA Elutin Elute twice t increase yield. Eluted DNA des nt perfrm well in dwnstream applicatins Residual ethanl cntaminatin Fllwing the Wash Step, dry the GD Clumn with additinal centrifugatin at full speed fr 5 minutes r incubate at 60 C fr 5 minutes. Genmic DNA was degraded Use fresh bld as lng strage may result in fragmentatin f genmic DNA. May

28 Ordering Infrmatin Prduct Name Cat. N. Package Size ABIOpure Genmic DNA bld/cell culture M501DP04 4 preps ABIOpure Genmic DNA bld/cell culture M501DP Prep ABIOpure Genmic DNA bld/cell culture M501DP Prep rd Drive SE Suite 150 Bthell, WA USA T: F: Fr Technical Supprt: supprt@alliancebi.cm May amplificatin prducts must be exclusively emplyed fr this purp

29 Made in USA FOR RESEARCH USE ONLY 2011 ALLIANCE BIO, all rights reserved.