Prot

Impoone ruone A) P

TinNnn

B) P1

2

C) PD12

D) W1

2

ocol for hy

ortant: Up toun, it is reco

Pre-heat hybTurn on the njection.

Note: Gettingeed to turn itot need to tu

Pre-hybridiz. prepare pr

100 ml re. Add abou

with the bfoil, put it

Pre-heat itemDuring pre-hy

. Turn on h

. Turn on 3injection a

Washing afte. Take the s

buffer III. While the

i) Fiii) Ad

sli

iii) Pr

ybridization

o 12 slides cmmended th

bridization smachine (H

g to set tempet on 6 h befourn the hea

zation re-hybridiza

ecipe: 74 ml ut 50 ml prebarcode at tht in a water b

ms ybridization

hyb oven to h heatblocks:and 65 C as

er pre-hybrslide out fromI for 5 min ee slides are sill the carriedjust the baides in the S

ress fluidics

n with MAV

can be hybridhat you separ

system Heat on/off)

erature and bore your hybting off unti

ation buffer (DI water, 25-hybridizatio

he top. Each bath and incu

, pre-heat thheat Gilson 98 C for dena heat plate

idization m the pre-hyeach. Use thoaking, prep

er of the MAalance. Set tlide Carrier.

s engageme

1 

AUI hybridiV1.3

dized in one rate your slid

and set at 4

being stable bridization. Iil there is no

(the same as 5 ml 20 X SSon buffer to jar can holdubate for 45

he following CP 100 tips naturing, 65 for sample i

ybridizationhe jars and slpare the MAAUI wash systo the numb

ent level ha

ization and

run. If you hdes into 2 gro

42 C at lea

are very slowf you run samone running

Tecan) SC, 1ml 10%a coplin jar

d up to 5 slid min at 42 C

items: at 65 C C to hold de

injection

buffer and wlide carrier fUI wash sysstem with D

ber which re

andle down

d wash syst

hybridize 12oups and wa

st 6 h befor

w for MAUImples serial

g samples the

% SDS and 0r and put thedes. Cover thC.

enatured sam

wash twice wfor manual wstem for spin

DI water. eflects the n

n and make

tem

2 slides on ash one by

re sample

. You may ly, you do e next day.

0.1 g BSA e slides in he jar with

mples until

with wash washing. n dry.

number of

sure that

 

3

E) MMthF1

23

4

5

flu

. After the w(full of DIthe lid and

MAUI mixerMake sure thehe Whoosh-D

For AO mixe. Insert the

. Remove t

. Remove tNOT touc

. Hold the First makthen slow

. Take the mMAUI mithe mixer

uidics interfa

wash buffer I water) and d start Spin D

r-slide asseme slide and tDuster to cleer microarray

the mixer frothe adhesivech the side wmixer adhes

ke sure the mwly lower the

mixer-slide aixer to ensurr.

ace comes ou

III rinses areplace the ca

Dry (80 sec,

mbly the mixer is ean them.

slide into A/

om the packae gasket rele

which will attsive side do

mixer is posie mixer to ad

assembly oure a good sea

2 

ut.

e finished (sarrier in the m, 500g, set al

clean. If the

/D jig with b

age. ease liner ustach to the s

own. Align aitioned corre

dhere it to the

ut and rub theal. Make sur

tep D 1), putmachine. Holready)

y have dust

barcode end

sing your finlide. as shown in ectly on the e slide.

e gasket brayre you don’t

t slides into told on for 10

or water dro

out.

ngers or for

the followinnon-barcod

yer over the press on the

the carrier sec, close

oplets, use

rceps. DO

ng figure. de end and

top of the e center of

 

6

F) PHmfo F(0thin T

F Ab

B

. Put the asImportanhybridizaon the sli

Prepare hybHybridizationmg/ml Salmoormamide c

or each slid0.1 pmol/ul)he volume-lnjection, so

The following

FormamideSSC SDS

Sperm DNAwater

Final volume

Alternatively,etter visualiz

FormamiSSCSDS

Sperm DNBromepheno

water Final volu

ssembly in thnt: Recordation systemide.

ridization sn solution coon sperm DNconcentratio

de, add 50 ul to your drielose duringthere is no

g recipe is bStock F

100% 20 X 10%

A 10 mg/ml 0

e

, you can adzation of buf

Stoide 100

20 10%

NA 10 mgol blue 0.5

ume

he hybridizat the barco

m and samp

olution ontains 45%NA. Dependon to get sat

l of hybridized labeled s

g denaturatneed to incr

ased on 45%Final conc.

45% 5 X

0.1% 0.1 mg/ml

dd bromephffer injectionck Final co

0% 45%X 5 X% 0.1%g/ml 0.1 mg% 0.1%

3 

tion system ode of theple name in

% of formamiding on samtisfying spot

zation solutiample. The

tion and exrease the vo

% of formam1 X 2 X22.5 4512.5 250.5 10.5 114 2850 100

henol blue (Bn (volumes ionc. 1 X

% 22.5X 12.5% 0.5 g/ml 0.5 % 10

4 50

to pre-heat te microarra

your noteb

ide, 5 X SSCmples, you ts number.

on and 2 ul volume is a

xtra volumeolume.

mide (volumeX 4 X

90 150 2 2

56 0 200

BPB) in hyin µL) 2 X 4 X45 90 25 50 1 2 1 2

20 40 8 16

100 200

the slides. ay, positionbook. DO N

C, 0.1% SDmay need

of universaalready adj

e needed fo

es in µL) 7 X 10 X

157.5 22587.5 1253.5 5 3.5 5 98 140

350 500

bridization b

7 X 10 157.5 2287.5 123.5 53.5 570 1028 40

350 50

n in the NOT label

DS and 0.1 to adjust

l standard justed for or sample

X 13 X 5 292.55 162.5

6.5 6.5

0 182 0 650

buffer for

X 13 X25 292.525 162.5

6.5 6.5

00 130 0 52

00 650

 

M

G) S12

3

H) SAwasTIfth

Make sure th

ample injec. Put the sa. Spin the

block set . Sample in

i) Taon

ii) Fibu

iii) Inmfothshappo

iv) Gfilwi

v) Lifinth

vi) Pucla

Note: Ketemperat

tart hybridAfter sample will check thessembly.

The hybridizaf possible (yhe beginnin

he sample is

ction amples (in 1.sample tubeto 65 C for s

njection ake the mixen the back ofill the pipettubbles shounsert the end ixer must b

orce to the tipe sample int

hould be finipplying forceort.

ently use a Kll or vent porill wick out ightly place ngers to prese seals remaut the assembamp. eep the lid ture stable. ization injection, c

e seal of the a

ation should you are ableg and the en

s well dissol

5 ml tube) ines briefly (asample injec

er-slide assemf the heatingte tip. MAKld be all aboof the pipet

e filled fromp to ensure to the fill poished withine until the h

Kimwipe to wrts. DO NOof the mixeran adhesive

ss both port sain in place.bly back on

of MAUI

lose the covassembly (st

be performee to see), mand of your h

4 

lved.

n the heatinga few secondction.

mbly out of tg block set atKE SURE tove the liqutte tip into th

m the non-taa tight seal.

ort. It shouldn several sechybridization

wipe off excT press too r. port seal ov

seals firmly

to the hybri

closed as

ver. Turn on tation seal te

ed overnightake sure thehybridizatio

g block set ads). Put the

the hybridizat 65 C. that no bubuid). he fill port oab end. AppWith a cont

d be quick (bconds. Leaven solution em

cess liquid th

much or th

ver the fill anAT THE SA

idization sys

much as p

mixing, andest). A Green

t (over 10 h)e bladders oon.

at 98 C for 3 e samples in

ation station

bbles in the

on the mixerply a slight dtinuous motibut not too qe the tip in pmerges from

hat remains ahe hybridizat

nd vent ports.AME TIME

tem and clo

possible to

d press B. Thn light indica

) of the mixer

min n the heat

and put it

e tip (any

r. The AO downward ion, inject quick) and place, still

m the vent

around the tion buffer

. Use your E to assure

se the bay

keep the

he system tes a good

r move at

 

Nte

I) P

2 ea

NEX J) M

Bu ON Up Le

Ri En

1

2

34

5

6

Note: Keep emperature

Prepare wasbottles of w

ach run. Ano

XT MORNIN

MAUI wash

uttons on theN/OFF: turnp/Down: moeft: move the

washinight: move tnter: perform

. Make surbuffer (Bo

. Turn on Zwill damaof the MA

. Select Uti

. Adjust baCarrier. Ttoo narrow

. Press fluiinterface

. Washing pFGA 06

Cspin-dry

Draw(He

Fill (s

the lid of.

h buffer I awash buffer I other is for r

NG

system sett

e wash systen on/off the ove the cursoe cursor to th

ng the cursor tom selection

re that all thottles 4 and ZerOzone. Tage Cy5. ZerAUI wash syility Menu/ M

alance. Set thThe carrier caw to insert a idics engagcomes out.

program

Cycle y (seconds) (seconds) at (C) seconds)

f MAUI c

and pre-warare needed. emoving the

ting (before

em: system. or up/downhe left, go to

the right, op

e lines have5 contain naThe ozone crOzone reducystem withinManual Primhe number wan hold up toslide. ement level

1

36 42 10

5 

losed as m

rm it in a waOne is for th

e MAUI mix

each washi

o upper level

pen the lid (h

e enough bufano-pure watconcentrationces ozone at

n 2 minutes.me which reflectso 11 slides. T

l handle do

2

20 Bypass

20

much as po

ater bath athe wash systexer.

ng)

l of the menu

hold)

ffer and the ter). n in Oklahom1000 ppb to

s the numbeThe track nea

own and ma

3

20 Bypass

20

ossible to

t 42 C em, at least 5

u, STOP butt

tubes are in

ma is high ao less than 5 p

r of slides inar the agitati

ake sure tha

90 sec

keep the

500 ml for

ton during

n the right

and ozone ppb inside

n the Slide ion rack is

at fluidics

 

7

K) R12

3

L) W1

2

AgitationB

BoBoBo

Recipes fohybridizat Note: It sand that you may

. Place the the agitati

Remove the . Turn off m. Add wash

disassemb

. Take the aGrasp thecoplin jar this procMAUI hy

Washing . Take all o

buffer I. PImportan

. Close the During we.g. (1) agat any timstarts spinImportanlift the dr

n (seconds) ottle

ottle 1 ottle 2 ottle 3 for wash bufftion station.

seems that iincreasing adjust wash

carrier (no sion will be g

mixer frommixing of theh buffer I (42bly basin.

assembly froe tab and sloin which the

cess. Repeat ybridization s

of the slides tPlace slides int: Barcode lid and start

washing, you gitation, (2)me except wn dry. nt: make surainage tub

40 Buffer 1

fer I, II and I

increasing tagitation ti

hing progra

slides) in thegood.

m the slides e MAUI hyb2 C) in a pip

om the hybriowly peel ofere is wash b

until all thsystem.

to the wash sinto slots of should be a

t the washingcan open th buffer inje

when it is he

ure the draine a little to h

6 

30 Buffer 2

Wash buWash buWash bu

III are the sam

the filling tiime helps t

am for your

e right posi

bridization sypettor basin (

idization sysff the mixer buffer I (42 Ce slides are

system. Fill the carrier.

at the bottomg program (Fe lid to mak

ection and (3eating. Also

nage contaihelp get wa

50 Buffer 3

uffer I uffer II ffer III me as those u

ime reducesto get slidessample.

ition. Make

ystem (pictured bel

stem. Immerfrom the sli

C). Avoid drready for w

the carrier w

m of the carFGA)

ke sure every3) drainageo the lid mu

iner is not tste buffer d

used with th

s the signal s clean. If n

sure the inje

low), which

rse it in washide. Put the

rying the slidwashing. Tu

with pre-warm

rrier.

ything is wor. You can op

ust be closed

too full and down if nece

e TECAN

intensity, necessary,

ection and

serve as a

h buffer I. slide in a

de during urn off the

med wash

rking fine, pen the lid d before it

you may essary.

7  

3. Remove the slides from the carrier after washing and scan.

M) Cleaning 1. After washing, put all the lines in nano-pure water and start Flush System. 2. Clean all the containers, buffer leaks or spills that occurred during the post

hybridization processing, hybridization and washing. The pre-hybridization buffer and wash buffer I contain a high concentration of salt and it will cause problems if left to dry.

3. Clean up the o-rings and the underside of the clamps on the hybridization system using a Kimwipe dampened with methanol. This may be done once a week.

For details of MAUI hybridization & wash system, please refer to User’s Guide provided by the company.