Amgen Inc. v. F. Hoffmann-LaRoche LTD et al

Filing 627

DECLARATION re #624 MOTION for Summary Judgment that Amgen is Estopped from Asserting Infringement Under the Doctrine of Equivalents of the Asserted Claims of the '698 and '868 Patents by Nicole A. Rizzo by F. Hoffmann-LaRoche LTD, Roche Diagnostics GmbH, Hoffmann LaRoche Inc.. (Attachments: #1 Exhibit 1, Part 1#2 Exhibit 1, Part 2#3 Exhibit 1, Part 3#4 Exhibit 1, Part 4#5 Exhibit 2, Part 1#6 Exhibit 2, Part 2#7 Exhibit 2, Part 3#8 Exhibit 2, Part 4#9 Exhibit 3#10 Exhibit 4#11 Exhibit 5#12 Exhibit 6#13 Exhibit 7#14 Exhibit 8#15 Exhibit 9#16 Exhibit 10#17 Exhibit 11#18 Exhibit 12#19 Exhibit 13#20 Exhibit 14#21 Exhibit 15)(Rizzo, Nicole)

Case 1:05-cv-12237-WGY

Document 627-4

Filed 07/03/2007

Page 1 of 15

EXHIBIT 1, Part 3

U.S. Patent

Aug. 15, 1995

Sheet 16 of 27

5,441,868

FIG.

III

1. AATTCTAGAACCATGAGGGTAATAATA
2. CCATTATTTTATTACCCTCATGGTTTCTAG

3. ATGGCTCCGCCGCGTCTGATCTGCGAC
.. CTCGAGTCGCAGATCAGACGCGGCGGAG

S. TCGAGAGTTCTGGAACGTTACCTGCTG

6. CTTCCAGCAGGTAACGTTCCAGAACT

.7. GAAGCTAAGAAGCTGAACATC
8. GTGGTGATGTTTTCAGCTTCTTTAG
9 . ACCACTGGTTGTGCTGAACACTGTTC

10. CAAG1~CAGTGTTCAGCACAACCA

1i. TTTGAiCGi~CATTACGGTACCG
12. GATCCGGTACCGTAATGTTTTCGTT

Case 1:05-cv-12237-WGY

Document 627-4

Filed 07/03/2007

Page 2 of 15

U.S. Patent

Aug. 15. 1995

Sheet 17 of 2j

5,441,868

FIG. 11

ETTCoaGIAACCATGAG GGTAATAA T~TGGCTCC GCCGCGTCTG c "T"ARI 10 AA
GATe TTTGGTACTC CCATTATTTT ATTAcnGAGG CGGCGCAGAC

£L~
3
,

ATCTGCGA~ CGAGAGTTCT GGAACGTTAC CTGCTGiA1G CTAAGAAGC TAGACGCTGA~GCTCAAGA CCTTGCAATG GACGACCTT~ GATTTCTTCG ,
.§

,5

7

TGAACATC ~.;£TGGTT

GTGCTGAACA CTGTTOTTTG AACGAACA ACTTTTGTAG TGGTGACCAA CACGACTTGT GACI.lGAA9 TTGCTTTTGT

.!

, 11

lt !
Q

10

!!~_IlHI TTACGGTACC G AATGCCATGG COTAG

KenI

II

Case 1:05-cv-12237-WGY

Document 627-4

Filed 07/03/2007

Page 3 of 15

U.s. Patent

Aug, 15, 1995

Sheet 18 of 27

5,441,868

FIG. 12

1.

AATTCGGTACCAGACACCAAGGT

2.
3.

GTTAACCTTGGTGTCTGGTACCG

TAACTTCTACGCTTGGAACGTAT
TTCCATACGTTTCCAAGCGTAGAA
GGAAGTTGGTCAACAAGCAGTTGAAGT

..
5. 6. 7. 8.

CCAACTTCAACTGCTTGTTGACCAAC
TTGGCAGGGTCTGGCACTGCTGAGCG

GCCTCGCTCAGCAGTGCCAGACCCTG
AGGCTGTACTGCGTGGCCAGGCA
GCAGTGCCTGGCCACGCAGTACA

9.
10.
1L.

CTGCTGGTAACTCCTCTCAGCCGT
TTCCCACGGCTGAGAGGAGTTTACCA
GGGAACCGCTGCAGCTGCATGTTGAC

12.

D.

",.
15.

GCTTTGTCAACATGCAGCTGCAGCGG

AAAGC~GTATCTGGCCTGAGATCTG

i6.

GATCCAGATCTCAGGCChGATACT

c .: vi .
FIG.

13

'ro i

EçoRI r;PlL:t Å ;i A ATTCGGTACC AGACACCAAG G~1\CTTCT ACGCTTGGAA 'CGTA~AA GCC;ATGG TCTGTGGTTC CAil,.'rT~GA TGCGUCCTT TGCATACCT~
;¡

'" "'

Case 1:05-cv-12237-WGY

.!

~ ;;

.!"

GTTGGTCAAC 1iì\GCi\GTTGA AGilTGGCAG GGTCTGGCAC TGCTGAGCct C1\ACCACTTG TTCGTCMCT TC1\C'fTC CCAGACCGTG ACGACTCaCT
§.

~

1

''" "

'"
vo

Document 627-4

!!

!' GGCTGT1\CTG CGTGGCC1\GG C~TGCTGGT j\1\7\CTCCTCT CAGCCG~ Cè-aC1\TGAC GC1\CCGGTCC GTG~CCA TTTGAGGAGA GTCGGCACCC

11

~ ~

~

BgIIII Bti.mI

l9.
"i,.~.

'" o ~

'0

-.
13

Filed 07/03/2007

1i1\CCGCTGC1\ GCTGCATGTT GA~AAGCAG TATCTGGCC'l GAGATCTG T1;GGCOACGT CGnCGT1\CllA CTGTTTCëtC ATAGACCGGA CTCT1\GACCTAC

l5

, 14

16

t. ,
oi
00
00 00

Page 4 of 15

Case 1:05-cv-12237-WGY

Document 627-4

Filed 07/03/2007

Page 5 of 15

U.S. Patent

Aug. 15, 1995

Sheet 20 of 27

5,441,868

FIG.

14

1. 2. 3.

GATCCAGATCTCTGACTACTCTGC
ACGCAGCAGAGTAGTCAGAGATCTG

TGCGTGCTCTGGGTGCACAGAAGAGG
GATAGCCTCTTTCTGTGCACCCAGAGC

l.
5.
6.

CTATCTCTCCGCCGGATGCTGCATCT
CAGCAGATCCAGCATCCGGCGGAGA

7. 8.

GCTGCACCGCTGCGTACCATCACTG
ATCAGCAGTGATGGTACGCAGCGGTG

..
10.

CTGATACCTTCCGCkAACTGTTTCG
ATACACGfu'ACAGTTTGCGGAAGGT

U. H.
13.

TGTATACTCTAACTTCCTGCGTGGTA
CAGTTTACCACGCAGGAAGTTAGAGT
AACTGAAACTGTATACTGGCGAAGC

H.
1.5.

GGCATGCTTCGCCAGT ATACAGTTT
ATGCCGTACTGGTGACCGCT1IA TÄG
TCGACTA TTAGCGGTCACCAGTAC

16.

Case 1:05-cv-12237-WGY

Document 627-4

Filed 07/03/2007

Page 6 of 15

U.S. Patent

Aug. 15, 1995

Sheet 21 ûf 27

5,441,868

FIG. 15

BamI BglII
GA TCCAGATC'CTG
GTCThGAGAC

1 5 ¿ ACTACTCTGC ~GCGTGCTCT GGGTGCACAG AAGACdbTA TCTCTCCGCC TGATGAGACG ACãCiGAGA CCCACGTGTC TTTCTCCGÃT' A,.GAGGCGG
Æ.

~
"-

GGATGCTGCA TC~CTGCAC CGC'GCGTAC CCTACGACGT AGACGÃCTG GCGACGCATG
j¡
1'

GCAAACTGTT TcclcTA'rAC TCTAACTTCC TGCGTGGT~ CGTTTGACAA AGCACATi-G AGATTGAAGG ACGCACCATT .:t!l J2

li

ÀCT~'\CTG
TGA . TTGAC

13

15
TATACTGGCG AAGckTG"~~ TACTGGTGAC CGCTAATAG ATATGACCGC TTCGTACGi: ATGACCACTG GCcATTATC I 1,6

sa..!
AGel'

II

Case 1:05-cv-12237-WGY

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Page 7 of 15

u.s. Patent

Aug. 15, 1995

Sheet 22 uf 27

5,441,868

FIG. 16

1.
2.

AATTCAAGCTTGGATAAGAGCT
GTGGAGCTCTTTTATCCAAGCTTG
CCACCAAGATTGATCTGTGACTC

3.

..
5.
G.

TCTCGAGTCACAGATCAATCTTG

GAGAGTTTTGGAAGATACTTGTTG

CTTCCAACAAGTATCTTTCCAAC
GAAGCTAAGAAGCTGAAßACATC
GTGGTGATGTTTTCAGCTTCTTTAG
ACCACTGGTTGTGCTGAACACTGTTC

7. 8. 9.
1.0.

C~~GAACAGTGTTCAGCACAACCA

ll.
12.

TTTGÑ'CGAACATTACGGTACCG
GATCCGGTACCGTfu'TGTTTTCGTT

Case 1:05-cv-12237-WGY

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Page 8 of 15

U.S. Patent

Aug. 15, 1995

Sheet 23 of 2ï

5,441,868

FIG. 17

EccRI RindIII!
AAT'CA AGCTTGGATA G TTCGAACCTAT
.~

3

AAGAG~CAACATTG ATCTGTGACT
TTTCTCGAGG T~TTCTAAC 1 TAGACACTGA

cPAG*'1"l'TT

GCTC 7\

GGAAGATAC T'GTTGQ~G CTAAGAAGC TGAACATC ~CCACTGGTT CCTTTCTATG AACAACCTT9 GATTTCTTCG ACTTTTGTAG TGGTG~CCAA i if ~

..

7

GTGCTGAACA CTGTTarTTG AACGAAAACA TTACGGTACC G CACGACTTGT GACAAGhAd TTGC'lTTTGT AATGCCATGG CCTAG

~

11

KpnI

BamI

-j ';1-

Case 1:05-cv-12237-WGY

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Page 9 of 15

U.S. Patent

Aug. 15, 1995

Slieet 24 of 27

5,441,868

lìG.18
1.
AATTCGGTACCAGÄCACCAAGGT

2. 3.

GTTAACCTTGGTGTCTGGTACCG

TAACTTCTACGCTTGGAACGTAT
TTCCATACGTTTCCAAGCGTAGAA
GGAAGTTGGTCAACAAGCAGTTGAAGT

..
5.
6.

CCAACTTCAACTGCTTGTTGACCAAC
TTGGCAAGGTTTGGCCTTGTTATCTG

7.
S.

GCTTCAGATAACAAGGCCAACCTTG
AAGCTGTTTTGAGAGGTGAAGCCT

,.
10.

AACAAGGCTTGACCTCTCAACA
TGTTGGTTAACTCTTCTCAACCATGGG

n.
12.

TGGTTCCCA TGGTTGAG~GAGTT1~CC
AACCATTGCAATTGCACGTCGAT

13.

H.
15.

CTTTATCGACGTGCAATTGCAA
AAAGCCGTCTCTGGTTTGAGATCTG
GATCCAGATCTCA1J~CCAGAGACGG

H.

Case 1:05-cv-12237-WGY

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Filed 07/03/2007

Page 10 of 15

U.s. Patent

Aug. 15. 1995

Sheet 25 of 27

5,441,868

FIG. 19

KtlnI
A ATTCGGTACC AGACACCAAG GCCÄTGG TCTGTGGTTC
L

EeoRI l

3

5
TGCGAACCTT TGCATACCT~ CAACCAGTTG TTCGACAACT

G~AACT'CT ACGCTTGGAA ACGTA~AA GTTGGTCAAC AAGCTGTTGA

CAA~GA

!

Ji.

AG*TGGCAA GGTTTGGCCT TGTTATCTdb AGCTGTT~TG AGAGGTCAAG TCAACcfTT CCAACCGGA ACAATAGAC~TC~CAAC TCTCCAGTTC
-s
I

1

10

CC'I§TTGGT TAACTCTTCT CAACCATGGG ~~gÇ~TTGCA ATTGCACGTC GGAACAMCA ATTGAGJ\GJi GTTGGTACCC TTGG~CGT TAACGTGCAG ,
1.£

,

l-l

¡ -*.l

! II

GlI~J:GCCG TCTCTGGTTT
,

.15

llI I .~EH I
G1\GJ\TCTG

CTIITTTdGGC AGAGACCAA:h CTCTAG1.CC1A G
,L6.

Case 1:05-cv-12237-WGY

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Filed 07/03/2007

Page 11 of 15

U.S. Patent

Aug. 15, 1995

Sheet 26 of 27

5,441,868

FIG.21l
1.

GATCCAGATCTTTGACTACTTTGTT

2.
3.

TCTCAACAAGTAGTCAAGATCTG
GAGAGCTTTGGGTGCTCAAGGAAG
ATGGCTTCCTTTTGAGCACCCAAGC
CCATTTCCCCACCAGACGCTGCTT
GCAGAAGCAGCGTCTGGTGGGGAA

..
S.

..
7.

CTGCCGCTCCATTGAGAACCATC
CAGTGATGGTTCTCAATGGAGCG

8.
9. 10.

ACTGCTGATACCTTCAGAAGTT
GAATAACTTTCTGAAGGTATCAG

1l.
12.

ATTCAGAGTTTACTCCAACTTCT

CTCAAGAAGTTGGAGTAACTCT
TGAGAGGTAATTGAAGTTGTACAC
ACCGGTGTACAACTTCAATTTACCT

13.

H.
15. 16.
17.
1. .

CGGTGAAGCCTGTAG~CTGGT
CTGlCACCAGTTCTACAGGCTTC
GACAGATAAGCCCGACTGATAA
GTTGTTA TCAGTCGGGCTTAT

19.

CAACAGTGTAGATGTA1~CAf\G

20.

TCGACTTTGTTACATCTACACT

Case 1:05-cv-12237-WGY

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Filed 07/03/2007

Page 12 of 15

U.S. Patent

Aug. 15, 1995

Sheet 27 or 2i

5,441,868

FIG. 21

GATe CAGATCTTXG ACTACTTTGT ~AGAGCTTT

DadI BglII i

G'ICTAGAAC TGATGAACA ACTC'lGAA
Z.

;¡

GGGTGCTCAA A1GGAAclcA TTTCCCCACC AGACGCTGCT 'ÙTGCCGCTC CCCACGAGTT TTCCTTC~GGGGTGG TCTGCGACGA AGACGPCGAG

..

.i

i ,§,
.?

CATTGAGAAC CAT~GC'l GM.'ACCTTCA GAAGT~':AGAGTTTAC GTAACTCTTG GT1\GTGAæJl CTATGG:AGT CTTTCAATAA ~CTCAATC ,
-s
,1_0.

1

11

JÆ.

AGGTTGAAGA ACTC~CCATT TAACTTCAAC ATGTGGCAA TTCGGACATC

TCCAACTTCT ~G;§AGGTA~ ATTGAAGTTG TACAC£GGTG AAGCCTGTAG

i ll !

i3

II

.l§

17 l' AACTGG~~~ A~ATAAGCCC GACTGATA-1Û'ttAGTGTAG TTGACCACTG TCTATTCCGG CTGACTATTC TTGTCACATC

Sal,l
1\TGTAACAAA G

TACATTGTTT ChGeT

3J

Case 1:05-cv-12237-WGY

Document 627-4

Filed 07/03/2007

Page 13 of 15

i
PRODucrION OF RECOMni!'A!'~f ERYTHROPOIETIN

5,441,868
2

oi"inree:iu:!:ciidei;a,,-:.!nor.C$,mc.ihcrormJlii);iol
a prOldn i, tlie iJ,iì:nate form Dr "i:~pr('s'¡on.' of the progi~mm~d g~r.(;1ic me~:sge piiwidcd by rh~ "'J;;!cniìdc-scaeC:DCt of"zcnc

Tilt, i, .1 CO(¡IÌ¡m~i¡()rl of m)' Cu.~nding U.S. pUnt

.'?f;ii~t)tt:'. DNA f.queii':"~ lisu::J!y .'prcn,dtO" ::
scn.e Íl ~ DN/i. pdymc( :i:~d plO'.-ide:i ~;¡e for ini¡ÜÜcn

al'plk::iío:) S~i, Nil. 675,293. f:ed N,)c-, 30, !n~ ;,:d issued ~~ Us. PaL No. 4,7ûl,WS iò¡¡ Oct. 27, 19r.7. whiet \.vas:: C()nÜ;iIi:iiion.in.prt of my copcrding U.s, p.:w'U ~pp¡k.ltkn$~r.":k 561.02:-:. fid Dc;;. I::" 1~~3,
5S2,lE, rtjd Fd,_ ii, 19$4, liilW :.bn,h:wd, :\n¡j :J
c-'iimi::\iilii.1¡i.i¡¡-p:irt or No. Set.

of ¡he tfJr;'-:1pliDn ìJlo mRNA. ..R"Oi1~¡oi.' I)~:A
~,:,,\l':_::'c~, .:~-c tm-,.~Hy "\lp~l(e"n:'" .of ¡i,,,.: p:"ççd~,ri¡: ¡;

gel\" m;, given DNA p()\ym~¡, b,m! protem,; ¡h~i (Jti::r-

flew ::bimdú:vrl, :i:d :i COiH¡f;Ur.ii()ri-¡:~'r~n of Set' Nc'_ F) min;: ili¡: frcqunKY (or ¡r,te) ofiran,;;:ri¡:lion:,¡ ~ni¡i;;¡¡u¡"

Ccikctivclv rdend to r,~ ":in;¡:T.O:er/r~;!-i""IOr". or
"coniioj" DNA -,e¡¡tlenc.:, thes" ~ea~¡ence~--whjch pre.
cede ~ i;di:c:ed 8"1li. ("r ~,,:-ic" d -",,;:,,$) ,,," !"m;I;O:¡3Î

655,S..i, fik¿ Scp. i,;,

!9~.: :iow:ib."COllN:.

f;\CKGRODND
The "iei",,! inv;:riimi rci.~t;:~ ¡;cncr:il.. ¡Oihe mantI',

DNA po!ymc; C(;pp"rillc to óetermine ,...herner the
15 tr,iiscnptiün (~nd ev"r.u,~! exp,,,:i,i,m) of;; ¡¡em: will

iibiiorl of gc:i.ilic rr;iteriÛs am( moie ;);H\¡~~ladY. tu
reço¡r.l;n~¡)t procedure,: m~k¡n¡; po,:siblr. ¡fIt prodU(l. iien o;¡ici),;ipt¡d~5 pos,;~s¡!'t; pn Dr :dl of ¡he ;:r:':\~ry

,.cnir_ D)lA K(;UCnCC; which "(ç,ilow'. " '.nc m :ò DNA polymn ~;d pro~'ì,!" :i '~tn:,¡ Ú)r renni,i:,ii,)t\ ö( ¡h~, ir.~I:,cripti0n in10 mRN.'\ "rc id"crefÍ l" .% ir~n.
~crirticr: "i"nr.i::ilcr.' "c'j1Jer,c",.

stfuCWr;¡! cQnf(,rm:ilior¡ 3nd/o; on,: or mNe of the:'O
hick!"ic::¡ p:opcri;~s of ,,;iiur;illy.ccccrrms ~ryll¡¡OI"':¡.

A foeii;; of miCHlh¡nlo;ìc-,,¡ pmc,,~'.il1~; fnr ilic J:,1
d"c:.J" !i:" b"",, ih~ "¡Ic"lp! W '''-'~nf~ct,-c ¡n¡justriall,.

and tih,mmccinie-lly ~iQ:iiiiç;:n~ ,\lb:i;m;:~~ Uôiiig 0(. g;,n¡;m~ which "iiher ¿¿'no! ;nit;~!ly h;wc g;:n::t!cßJ1y A. Manipulatioii Of Genetic ~!atcd,,¡' cmkd informaÜon (cnc"w;ng 'lh" jn:i-d pioJuÇ¡ inGciicii,~ matenal; may be broadly defir,~d a~ (:'0''' 2S i:u(!d in ihcii- DNA, Dr (in ihcç;..~~ Qrm:i¡¡r,~li;,ri c('js el,emjç:ii "Jbi:m:i: w!Úch progr:im fer "iit! gt:ic" ¡he. in culture) do nOl \x¡Jin~rijy exprd;~ :i c;iiom(;,omal m:iii\lf;¡çt\lr", of co:istÎ~i.ents ofcdl$ and vini,e, :,rid gene al :ippreci;hk ,evds,. Simply pm, ~l t:cm: rÍ1:i1 ~pe:¡. direct :hcresooscofccl!sand vìr;isc:. A long di~in fier- ~hc ~miciure d ,¡ d(""rcd pnlyp"piidc product i, pülyrreiic s:ib,l:mc~ known æ¡ deoxyribiml!cr.;c :,C¡(i eíiher l~ol;¡teC irom:i '.,~cncr.' org::im,m or cb:rric:.ly
t"iJr._

(DNA, compos... the gcn.:tic m~t~i~a¡ "ralì Ii..'ing cdi~

:;0 ~)'n¡hcsíi.ed ~:i~, :h"n, si;,bly inir?¿i:ecd into :;~oi¡':cr

;md vin,:sc$ except for ceruin viru~s which ~m" pra. gn~1ned hy rihonudc;e ¡ic.'¡d~ (RNA, The rq)'''lirg i:niis i:i DN..\ p:'iymcrs uc four ¿iffnent nuc!cotide;,
each 0f whi~h comu.L. of C;:hCi ;l p\ll;"~ ("okn;"c U(
gu;m¡~:e\ or:: ¡:ytim~din" (lll~'nÜ:i~ or çy\u~ia~) oo\,~;d to :;

ùri::ii\'~;m ","!ii;:n ~s prdcr;ih¡y 3 ,di.¡"ph:"l\l\g ;)l,cd,n_ iH org~ni~m ~.ueh ~," b"~!cri~. yc,,~¡ N m~::\m"\i~il çdh in c\.ln,ii:. O:"c:: ihi!. i, d0~C, rb" i:Ùjì:i~ i¡i;ichir:ery for :.ell~ ",'ÇJ""~¡"n ¡n iliç .'n~liiJ()::-ne;!.' 0: .'ir;on~(.;ctt:!"

mi,:wbi:: hest cd!:; crnat~;; to cc";nw¡;ct ih~ (kÜlcë
p:OdtJ_~l. 1,~in3 ih~ c~ol;tn()ll. lJN,\ ,,~ ;, lcrnpbu fDr

;; (ko~yrilA"",,, \UR;' \(l which;i ¡:hotpli;,tr. grouF ii.
:11:,d'ej, ,\ u~çhmcnl of nuckoiidt~:n ijn(!~r FoJym"ric form is by mc~¡\'i cffu,:ui of :l\t' 5. r:\Ç:.i¡i)).)rc ')(en" nuci~()ii.:o :0 Ú:c j' hyóro~y¡ ¡tw"p of ,,¡¡,,¡hcr, Fune
~.~f;Wij,,,¡ "",:.d"i;on,; of $ingli: ,;ir;:n:l, çr m;c;ccû(ides
(I:MJ',_n ~l dco;ya¡igollJci:o:i:.ks), \i,beli ;:,~,(Y~'~;i1i,:,p'"

ir:i"::ripiwn or mKN..~ 'If:di i: Ümi ~~3ri~,bir.d ini,,, :\
"0"lmU"u~ ~e¡¡uc:;ec oj ::mtn" ,~::,d ':;_",'"l)C''

iioM) DNA (,)(cun in the ibrm of ~l"ii¡" doubl" ~(! rd~iìi,j" ~o '.reWlt.I!;;llant r)N.:\ '" mc:hcdo!¿:;:!"-~ (('i ih::
Isol;iUC;:;, syaihc,i.s, puni,c_;-i,''', ;,n,) ;1:npliJ¡c:::llG:l ü( gene-ti;. 1l:1¡,,ri~!~ i~;r \1,," ifi :hc ¡r::Il;hrrr::¡l;C-n of ':e_"
)ccicd )¡0sl Mg;;~ism,;. u.s. P"t "'G, 4,2.ì7.::~ wei¡.
lWll. n ,,1.. :.", n",~i'pk, r"I;¡~, rr, io("io",.,,~tj,_),., c,i-

The ~~¡ ¡~ :.ich '" i'~:"i\~ ;;:\(i );"'"'1''''' c"b¡¡'-"tiu~s

L.;:.ur by li':Jm ()( liy~rU'si::: ooiuEns l;,¡.,i.t.~n purint
;Ifd pyr~m~,~m" b;i\C ¡l,e.. o.:::mpkmrai;iry :L'C"C;~.

¡kms e~isÜng cithc~ ;x'lw~c' ;,dcllÍ1\e (,\) ,mr: ibymir¡c ~j IT) (\~ i:\l;I'i1i~e (G) ~i:d cy:o,im: (ei), By C():~vi:nii(ll. m,d,'üt~(k.' "'" ~d"neO: I" t,y 11:" n~m"" d ~l."" ,:c\",;',iiUt¡¡l !,\jn~'.~ ,)r ?,'¡ir;iòi11; \;;i;,"\ ;,.'d ~hç çgiip!cnw~i.
t"" .".,~_;-~ :,¡'0n" c1i llll;:"'0~ld", If ,:mJbk s;l';imk,¡

uii~ccllui,tr hc,~ ';\~.i~n¡:;m, w;'ll '.h::h',J" virc,i n, ç;r,-.".

);;r rl~'i;-,d ut"!, "..riidi iiidiidc:i .'ddi",d "we"'")""
DNA ":q"c~i:,,_i;. TI'j(. p~G"",i1l~" ,,r ihe- COLC'1, ei "i
?~¡"f;t fir,;: !~i'.-,)ht i;n::'¿;,durc ,~f .1 (,,,ni;r,-i~m.1:;''''\

'..Çct01 bv i:llt)1J~"ti~;,:!\' d",,;,;,I~ ;':L;l ,:i:-""i"r pb,;:"i,l

DS.., ,:i,, ...T ~nd G-C) :.:c ;d¡:n~d 1,) "" ..b~,c ;,'.ì
""" '. ' .,,,-'~',~ "',,. ",.~ '::':.~ ._C" , ".:'. ":' ~ '~.."~,...~~, - ,.."';d ,', õ "~¡\"..'.'I'''':-¡_ ~'..'i.-;";,,. ~

DN,-.. u him ;¡"o;:; D¡-.A ,Il;m;ic ~tk.;il.',j !Örcitm
:';;~'~:¿¡;i~\.~~~i:~l:(.:~~ ~,~:i,:;:;;)(~:¡")!~~;~) 7;;~';~\"~;tr ;,;~':,~ ,~~:i¡)~;~~

.,,"C", "", ;: "" n "'~, ç y:o,~"~ ;,"'; ',;r ¡w:! "..). ,,,i¡'n : r;: n

ti¡~~i~~~;;,:~\.¡¡\:~~'(:~!~~;,:.: ~:;:~"';T('.ti:, 7:~L~¡:':~':;;~~i '.:~'~r~;:~;.;'~

::¡ ;~i:,~,~¡~':~;~~::~;~'::~:;~;?~~~~~~,;: ;~!~~~!~~:;~i;~£~~~~l;':~~~~ ._.

:'oi"í2D D~A i~, ü,,, rl,,~,,'c,~,,!) í~s.~:in~ (;n?:,'~iC;, i:,,:,~1;,c ôr "iTccii:~2_;' r::,\(;I';:¡i:'n p:çC:C$:, '''1:1 '.i:'.-hriJ" '.eç.-

D~..': r::,r!,,~,:ide ,.cau"l:cr:~ (£,'nt~) ',:" ..Ir~';'-:;:t,h:d'.
¡"i(' I'd;J"'-~'y 'i~s,,\;¡c r,~",","¡:~r ¡.l"", (ml-~N.'\:' F-C)!\,.

i:oC':,_ Tk 1,..R~.:\, in 1\¡~:~. :,,,ivu, ;,~, ~ (cn;,.!.;i" k,:k
fi):m"ii,)i\ .::f ~,1r;'~:ë::;;t rc;:\ll~(Ory ;':-:¡ ç;,~::¡yt;c l""'¡r ¡", rw,,-, ~"'-¡;n,:¡,,¡:i;;~. Tbi" mRNA ntl""I."L,\i.:i:.. f",-~C'" (/)

..'W, "rt f0''''"'' ",',,,;oi, ind ~(k ¡);,; ;;"),-c~,,,j '.M)~,~'W;:' iJ:~A ~,':;'n~,.!~i '.,pi c"d ",:(, ~h~. -'.'n;_,: (-_r ç,¡"-_"L,, 0'-,\
-";'~\1¡;U

'¡ .¡"~,,Jc;, :;--:,i.:"" ,-,f i;,,,i-,!,,¡ih\" to:;,, d111';'¡ ~:,)" or¡~::r;.

:~".'(;:"':' ii,,' i.':,~r::\i..-,~" d ,ird! RN.. '~r",nd' ,:R. ~''''.;
W~'h,')' ti:"-''--Vn-1 :;r.'~ ""¡:n '~'1,vi,ii::¡! ;,n,:-,' :,r;d" ~in1!

',rm'..-,ihik
''''''(;rh'

h;'~,,,d \"~~..:,)i :'i:::_,,!l; ,,-, ihe- f,.:-_;,,-,;,!¡c,,~ ;~!

':'1 ir,,, """,'''''''''' j)~,.\ ,~; :ii", ;:c,c'~ c'c,'
¡omc ~r,' :;:ncc-'" ¡!-,~ Ô;:,i:-..d l~':,"¡: ::' ~,ir¡:-

i;',,~ mr'-~:\ ':;~~,nd ~ù ~h"\ !N ior:n"i,r;¡ c,:' ;o0;:.T~P.
W;c-S ::, Fr::'-pcr .,:¡¡¡r0 ~,ç,d i.ca~ci)(t;".' k ",H.:....'. ""1=,:.

;:.,,:;.;i,\:w::

::~g~ c:c.ri'.'d iw~, DNA. :¡"Ô Fr",'icti"f :b: k,."': ~'i'" Ù) n", If(!\,\ '.J?piy ;mc or:~nt~ticri of ~:\J. "i..'."" On" "i¡hc:'.'cr;:"",'fi¡nü:;çiJ~fnrl!ü!yrcpi¡dc'.c.\pc,,-i()l... ¡,_

¡:;y,L"",n¡i;:("'.'i:(!'l,,i-ulc.
!¡M','cs¡c,;i,cDN..\ ":Jk

D:\.- ;wi. .ii(

. \h~ F')~! 1i~~,f:)"ri':l¡ljn ~:: the ~~pt;,5~;:I~ :'y :1-,'' h',');i ,-.c;!\ ",(
¡jie "xo¡:¡,no\;$ rX';A in :h~ (()rm u: br,-;t ~.Ç:;k ~.ymhr_';:';

i:, '!I' !();:;I Dr ¡¡ipk: '.co:::mç' -sc;:t;~:i:¡:il gw\ipiq:,

(J:. ì'-(,hi;;b:t" (,U;¡niiW" of :;,\:!\n:~rÓ;,¡¡y- ~\Eni¡",cè_nt po.

Case 1:05-cv-12237-WGY

Document 627-4

Filed 07/03/2007

Page 14 of 15

5,441,868
3 ieir.Grr",\¡i'l'~f'tide(i~¡;:ni:nl$cxedf(lri.ythcfc=gl.
DNA. $e~abc, e.g.,

4
the di5c1c:l.re :.nd ti~\l\s¡om of the ~rt ¡:mvìt!e(l in

4,2iUm (to M~;Ú). ~,2-)3,652 (to Celie-A). and Euro.
p,,:in P:iicnt App1iq¡tiO~1 093.619, ptiolisheè No,', 9,

1953. .

US 1';,1. Ncs. ,l,2c-.,ni (to$1i,n",J,

U,S_ Pal. No. 4,394,443 10 Wei~m~:i, ei ;11 Jr:c inc recent d~:-c,i:~ir~iion~ "f the = of long. oligo:\ucle(l. tide hybridbiion proè~ reponed in W;¡J~;:e. ct ii1.,
N:.c. .,lcf); Res. 6, Pl'. 3543-3557 (1979). ~,nd Reyes,.:1

The dcvdo¡ìmcnt ()( .ipecific DNA $eai""oce$ for

~L P.NAS (U.S.A.), 7$, Pl" 1270_3274 (1%1), Mid
hye-. e-I ~l., ,".'ltc. AddJ Rt~, II, pp, 1315-2335 (!'ìBì. Set: ;ilw, US POlL. No_ 4,35£,53510 F~;~()\v, e: "i., rcb:.

$pHdne ¡nie: DNA \.eel()~ is :icc0mpl¡~h"d b)' a i,a~et).

H);) gr""ict:¡ on :hi:di:grce of "forejEnne~'" of ¡he "donor" io the pwjected ho,;:
oficchniql1";, depending

ing to DNA/DNA hybridiztion ¡:t1ccdure~ ¡n dre;¡"
10 ing dbsnö~i¡; publhhed Europc:.!l I'~I"rI: Apr1;c.¡(;O"

aiid :he $iie O~. (he pcly¡:",ptid" lû be c.Jprc~\cd ¡n the

lw,l. At il)e rìÚ: of '1"c¡.;ìmpW\:;ilÌcr., it "'lfl De ,t;¡icd

il'.:: :hrcc :iltern;i:ivc pindpil mcthlXs c:m be em.
ployn.L (I) th;: "i~la!ÌQ~I" of ûDubk.str:indcû DNA
~eq\lcncc rrom the iic1iciiic DNA of ¡he ct01\O~; (2) (he

XO~. OO?06~5 ;iiid C(¡iQ6f rcliiiiig to lii;h:--mìtliiig
íiibch ün ~ing!c 51nndd polynride(;tide pmbK; D:w¡~.

et ;)L "A MiinU;¡! for G~n"ik Ene:iii"cnng. Adv:.ncd
ß:!c!c:rial Gel1clic~H O;)ld Spring Ihrooi L:ibc:::ory,

cl:"m¡ç;l11~¡n¡f:Kllrc ùf:i DN" "".,¡ueuc.: rf(".'jd;~8 ~ b Cold Spring Harbor, N.Y. (1%0) at pp. $5-% ~nd code ((\l " .,¡jlypcpti(k ()ri;¡i,,,~,i; ,mll (3) ií'k in viirc l74-i7Ii, ~ehiing 10 COk~ly and lb~u" hybrici~.al;(j:l ,:,'nihr..\i~ (if~, è()llbh~_,tr.¡;dr. DNA ~c:au,~c by elitS' :eclmiql1L';; :ud. New En¡;l::md Nui:le;lr (¡k)~lún, i--fii$:S.) m~iie "rcveru: 1r~::!;r.piicn" Df inRNA i~(lhlcd from lmichures for "Ocne SCTc~n" llybridiz.:iioii Tf3.11:Ji:r donor cd!$. l1:e l,~$t':nen¡¡oned mcihod~, which involvi: Mcmbr:lle m.itcriah providing ¡i;~lrucr¡(m m"nu3h Îm formiion or:¡ D:\A "ccmp!i:r.êm" of mRNA Me ¡;C:1' 20 the tfan5fer ~iid li"br;dìi..li'¡:1 of DNA and RNA, Cila-

cr:lly rderr~,J 10 as "cDSA" m~ihoÒ,
MJn:.f,1Ci\lr~ or DNA £~qu~ncc; is rr('aiicll1y the meihil~ or choice when ihe e,ltire ,c'l""iiç;; Qf :miiHu
:'lâLl H,,,i'¡uc, or ihe deli

log No. ~EF-9Î2,)
Among the rion: ~ignjfic:mi recent idv:!t:ee£. in tiybrid;i;itÌ(\npro.:ed1Jre.~ftlrrh"':trttn¡n¡;ufre(''imhinanl donC:$ ì, i!i¡:u~e cfhbdl-:.j mi;i'd $;'tiihetic ohgonudemtion simple ir.dudîng ~ !:eicrD.&C10Ul m;~i,"rc of osn.
tic s:riuded 0:-,., Of RNA~. Tlc~c pr,oced(,ro arc

red l'tiiyp"pii'k produci is

known DNA m~rilif:ici\Jrin3 l'ic"'cdme, of co-owned, ", c!Íde- picbes, ~;1ch or which i, ¡iotemi:1l1y the ccmpklt co-pending U.S. pJI~nt Jl'p!k:iticn Ser. No. .H:;,'UI, by complem~iit of a ~¡xci¡¡:c DNA tC:lltnei: in ih(. hybrii.
Alton, d aL., (liled Apr, IS, 19~:i ~iid ('orre'¡lY.lndirig to

l'CT l!SE:FfJ605, putli~hcd ",ov. 24, 1983 ~,
vidin,; for \hC-l're~li:;e()rallernilee(Xjmi~eominoniy
found in ¡:ionc\ which ;¡IC higlily e.ipfCS.l¡:C iii tiw ho;t

WOS3l(l~O$3), for e~~.mp1c. provide II ~"'pcfior m~~n, ~chiowkdgeJ w be ""1x:,.Ù!ly ,,,e-ul ;ni),(: ùeie"I;'.m üf for ~cç0m¡;I:;;hing mch hi¡thly (ksiubl~ rewhs .t.\; pro- Ju l:DKA doi\;,: dcrivd from WIl!e", which pwvjil"

c~m:mdy kw ~mo-tllll~ of iiRNA uaucr:CI' flj ih"
p-lypcpiid:: of ¡nletc,!. Briefly put. ,"sc of ~:rineenl

Ofginismsekci¡:dfori;xpr""j()n(c.g..rrovidinß:'=xi
or E.r:;f "l'rdr,reni'"'' cr"kn,j: ~w',;ding 1b" ?f')'(el)Ce

hybridiz~iic~ cond;lioM di!ccied i()wr,¡d ~vo¡daiicc of

noii.~pccilÌc binding C~n "Ikiw. e'$.. for 1...:c mJior:.dio-

i)cnir;¡n.lbli;;J "¡nlron" $equr:ncc.l (commonly rp!::t"i ;~ i\r:ipliie VìS1~¡iz,i(i(jr. of:i ;:cÒfic cDNA clc,¡:c upori ihe in numm;¡1i;¡n genomic DNA ,(:q~ene,,~ and mRNA ,","ciilofh:,'bridiZ2(;oriof!het'lrr,etDr-',\iO::UISi,I!,,lc tr~i",crirl.l t~(:f(rd) wh,eh ::re nN rc::dily procesg~ by probe within the mixiuit ,,,'hid\ LS ib ¡;Oiiple:" ç.ifl¡ilcprnc"ryoiicho.licdh;e':úi::ingexprc."i(llioflJn:ie~ÙtJî lii"!'C S"''' grllc:¡~lh. W~JI;;e. el ,,1.. X::c. AäÒ P,,'l., 9. "!e"dcrH polyrcpiidc sequerice.lc0.mmc,l,!Y coded fo! Pl'- S-i9-Sn ona SCßgi-, (;1 ;',1. I'.N...,5. (C.$,.A.). n. by ¡,enümie DNA ;ind (;DN"..I" ;"'"el,;:o: bUi Í¡"O(lu"a,ly ~'- pp. 66!3~66l7 (931); ct,(lCi. d ;,1. Net,-"". 299, Pl'. nO! re~d¡¡y de;;\'cd f,oiiillie pu¡.;'l'epiiJ:: ()finteio:: by 17~-iW (1%2): Ki:iadii, el ~L. P.N...S (U.S,A,). ì~. bJ"(~:':"¡ lH Y~'lsi ¡',)~i ecH,: lf'widirig for ("",dy ;:\;.1. pp. f,.¡61_(~\e.. 098l); ÚI,kuD0, e: ,.:., ¡'.N.AS. (C.~,A.). ;¡on of ÚH, DNA in convenicn: exp~t-:,;01l vc"iç'l$ in W, pl'.:! 196.12:;) (! n:;): :.iid K01n1;lih:i, d ~L P.,vA.S.
;;~;N;::¡li:\:1 ,,,ii!: dc¡¡~c¡j pl0:!,Cle;!rç¡wbior :,1);; term:,

(U.S.A.), BÜ. Pl" 321S~3n? (1923), ír. ge::c~.,¡, ihe ii:IOIicaii,'r,CC::?l1dp;(wij¡r.rf.orrc.:dyccw,tr\;c,ti:in ~, mixed Fwb.: pmced\lrc: ofWalb:e, c(.1. (i9~1), ,\jpr~.
¡wsc hC~'1 "~p.¡iid~d \lpO' by C-.;i;o-ll' w0r;;cr, (0 the
"fi):" (:o,;¡¡td 1".iiYP"l'ii'¡cs . _ .. '. .

D! gene,; CCCli;: :01 polypq;ti"" t~"f'm~r:u, :md "n~W¡;~

\\~:C¡¡ ih~ i:r.llrc $c'liicni:c ,,! .1ml"c ,',,:,c rc;im,,,¡ d
the .;~:i"c;~ i,,-,I~p,?r;J~ '.' ,,,)I b0~'''':' ¡¡,n:e! i;',;,;wr:;çHU,' :',: l)l\A ""'-"':'."'" .'_' ¡wt p;',-~¡vk ;~",¡ t".):;,~wn.~f

rú;n: wlrocre ,d;"bk :c~uii, ilJve .."poiic-dly b",,,, lib. i;:.:I,;J if: ~ ",DNA d",m: ¡,obi;!):, ,,,ing ;¡ 3~.n"~f!b," m;~ed "reol" üf i,;"lu;c.lDnf: (lti.m"r,0liRomJd~oiid,~ w rr0b'~; of Hn,(onl1IY, l';,r;.'1S UNA ,cs,,~n"t.~ !igeikr
wiil¡ ~,sinf.k ri.rrer to dtec¡ ~ ¡we.

DN,.. ;,'!ucnçt;. ,'()dm~ i,)r th~ ¡:01ypl.,.U:!" ~,\, _, "D\" mdli~d bec'\r1\:'-'_ :b' 'õ\e(1nÔ c,( d10ic',_' ,k'ri~~ :"" pc_
¡~:i:j;\i dr';i",h;,,~¡;'; i:'. ~:,~" () ;¡¡,emb!y (,~' ':XF2i';l(ìC1

me "p",,¡Ü"e." ::00-

f;ITutioli ¡)l th" plc,-"ce çf cDNA d" ir.i~:"q. S~".
Singtr.S;,n'. el .,!, ;-_N..I.S. (U.S,A.), 'm, Pl" 50;: Fro
(19R3),

vccwr~; c~¡nhl~ ol p',w:(!w.iò !Ùt" 1",--,,:$ ()f m,ç¡".hd
(:.~l"";':,:Ui ~C.k~1cd!(. ;i""'.'''_ Ar.~-ni.~ t,." q"",i:d p"'-' ."

~cd~~c, íi:r çDl"A """l"'~""c'o i_'( inwrnt ,s ih~
prcp,,~~,i¡,;n .)r ph,md.h1:'''c ~-DN:\ "¡¡'-r"ric:' dc:i',,~;!
hçin ,';",'(,C ir;,n¡;:ri?Ü¡¡n d' fi1IZ~..\ ;,i;u",Lm: ¡¡; c:,-mui cclll ;c:e~:~rli~, r"sp'-lfÔbk i,':,¡ bgh !n'd ""Trc,'~,~()n i.:l

Tho: "" '" ¡(ei"_":';", Dl~,,- i;~.b,"ô;; '.hoc ic;,;, ,"::m. nwn ofiLc tLice Jbü\'c:.n,:i,,¡j nielÌhJJc for i:"v~bi'il1g
:'.J",;,ifi~,?N__\ ò\:t.i:c:ri;.Co fur 1.l''~ in fcc"rr.ti1\:l:"t i.-,nwf:' ui'r,,~'. i hE i\ e,p~Cl:iJi,. ir',~1: ir, ¡b: ,i"':~ ct i-i:,~~,rr()¡~'''1

¡:""c., k,t,. ¡;b,'''¡~' ,-',f d)l".-\ Jc:',,-,'J t~"rn ,UiU;:"I;.'

pn::ccdcresdircc:tdW,ecHrinfr;ic,(;.hi"lnp:'c"sicnoí
~'~'ii~:~:~:i;;: ;~~~'~,~(~;~~n~;::; ~,~~~~~.¡ ~ ¡~:~;¡~. ;;:,;;~' -1~~;;~:

,':, m:""",.1lio" r__nly!"'pl"~'6 ",,¡ ¡, :'",'- pr.nc;rl'¡y ie :hi:
¡,hr~ri:),; of¡:enornie D~/.. ",i"f:'m,,:i ;,r,d ,,:h~r m~ri~;r;;,. I;:," :',r,cçÍi.'o ,-'i~gìii, 1St.-, t.!;.. Li\';:\. ct ;ii C~.'¡. !~ pp,

eel!, \\!ad¡ oP""_'.' ~d"I;,"\y !;,r~" "i,;".l::;" of ,2¡"....."i

)¡..-t:;¡""" ¡",,,i,,,'.i,:' \'.ht'" q!.~t:i"l,:d 1''-1';:''" ,..
Viìyp,:pi"k', .imi"" ;,,",j ".-;,';~';~~ :ac' i:~.,!W'" bbdic~.
,m.;~k.,:r;",;,d D,~..I" "nile ''::;\:c::;c,._; ¿"p\¡G,,!¡,.,~t "

~~a~~~;~¡~!:~;:;Jõt~,"g~: ~.~ ,~~;~I(~"¡: ~:¡~;:~~';:~':'~~:;, "~~ ~.~~ ~;~:;:
ç:.rrl"d 0'-1 on d"n~i! "~Ti", nf 11:" cDN.-1" which h:",,~

'" ~ 1 ~.i~_l,¡ ~~, (,¡~;,::\.. rd",.i:~,¡; id~ ,¡:,l~;:~~i,Ut:~:~, :ç:i. yrre'~l:~'~

V;LIl den;l~lHd lu oili¡;k ,(i::~"b:! rurm. ¡S"I:, r.tncr~!ly.

~ ¡,iin'_f. ~c.~)...,~ li\J....~ """",'''.,., ","i,~¡J HJ .,_ ..1e "l'hni;His !.br;)iy": K~m_ ~l ~L. f'.,\~A S. (USA). n, PI'_ 5172-51;6 (!9St!) rthti:ig !e' tlllim~¡; ;Jltnainic Ji.

Case 1:05-cv-12237-WGY

Document 627-4

Filed 07/03/2007

Page 15 of 15

5,441,&6S

5
brx'. b~,d on ::!icmativc rc;:o;;ÜQ¡) cndo:iuck;iSC

6
ihrcc fom!s: i'. /1 :m\i ;i,,\~io. The a 41':C ii f0r~;1' difi,'r ~li¡1htt:. in c;ibohydr:tc comv(lr.~nt.t. hm h~\': ih,- ~;imc po:ciicy. f;Q10gic:.;! .,(,ii,',!;, ;ind mQk~u¡~r '.vc;ght. T:-,~
hi:,li) ferm is ~n a or ,': fc('m with ih" l"rm¡¡¡~1 c:;r;;üh..,

Îlagmc¡¡I;¡tioa proccdu.rc; am! BIattm:r. ct aL, $',,'('11,,\

¡96, pr. ¡61-- iS9 (197,) d::~crib:r:g Ccri~¡roti(Jl oi ::

lxwinc gtnoir.ic libraryl there have bee:i rel:iivdy t"w
~\ièc"",sÎ\.il a¡¡cmpts..t ure(lfhyhr¡dii:i~io!l Fo;;cdures

dr:Hc (,iilíc ~;;id) n:movd. Erythrc;:0ic!ìn is rrc"cni in
I'~ry low cCi~ci::;t;'licr¡$ in pl:in:~ wí~,,:~ :b; bel" ¡~, in ~ hC;'llhv ;;iiie wbnd:i lÍ~,~ui: lCr,~;Ye 'iJ(¡r"i"", ()\Y~C:"

in ;~(.bii:ig Ecr.0mic DKA in ¡h :lt)$:r.cc of c.\knsi,'c forcb:ow!cd?,c Df ;¡:nino add or DNA ~qticnces. A,
oae c~ampk. F¡dd~, el :it, J. MÛ. ¡;r.¿ App. G",'NÙ:s, i.

I1"lion f~om the i;,\i,i¡llf¡ m¡mbn of cryi:\!CC)1CS. Tht

;ir- 3-18 0%1) ,eport thouccc..-ful b;)bti0i) ola gene
beman p¡!\iit.~ry gi~'G)' i0 pm:";:; hormonc~ frum ihe M:ini~ii\ Lil)~:uy ¡!:n.)\.5h
coding Îw ¡he alpha ~ubu¡)¡i of

¡'¡:m:" j(,w cC'f:cttir:itioli i5 et:cmgh ¡c, srimiil~:" i",. phc,c\nen! of red bbc.;j cd¡~ wb:h ~,r~ 10:1 norml,lly

i.seof:¡ufuiJkngtliuFobinc1udingacomplct,,6?1
b~e pit fragment of:: previoudy-¡"ib:cc cD~..\. 'c.
qutiiccfcrt!ica!ph:i5\biiiit. ,\~;i¡¡o¡hcrc¡;amp1c Daò,
,,1 aL,P,Y.A.S. (U.sA.), 80, ?p. 1531-1535 (19Kì) n::¡Klr~ 1:

ihrc;ighigíng. Tht ~:nriunt ",( cry:hrccoi~tin in :ht ,,¡~~\:J:tion "
in¡;r";,,,d under cD::di:,cl.; of hyrQ~i" whcn oi;y¡;~n ~r~M¡ion by bbod cdl, irl ihe circ¡¡!:.!Ìol1 i\ ¡"(h:a,,j

l1ypi;¡~ n"y he c~ustJ by k~~ of big" ;òi;oum, d
hb\)d ih:üugh h",n;()ih~gi:, úe~!f\clÌon of ~~íi b)ot;d
cdb by oV"T-e;ip()~Ure 10 r~di:.i;a~. r",dUC1¡,~:i in eXYIT.:n

¡_\(¡!:ii"" t¡flmnian ¡'",ntimk ~'.I0n:;,f':H h~lm"n HL/..DR

a,ir¡g,l 175 b..,-'e p;;¡~ ,,'''lht,c ùliglillC;;!~úl¡Jc. Fiidly,
An(.ki\"". c: ai.. P.NA.S. (U.S.A.), SD. rp. (,S:S-6$~;

;llLih lÌi;~ 1." hi~h ;;I:im~(", or prolonged uiit:or;,;:ious.
:,r_s.;. "r V;!!C'U~ turm~ 01 i::emi:i, In n:-~.?c\¡;,C 10 t:"r,',i~,;

(19S3ì !-ipen the ho1!lÍon nr geilOmì;: "lon" fM lx,~'i:ie

Pf);:R:,¡¡" rryp,ir, inhibiWf (BPTJ) uti¡;g ~ single pw:i ,ü

\)nrkfgoi~g hypexic stn~;~, cryihwpoìelin w,n in¡':rc.~~~

86b;il:;r-li¡"i:ik!lgthandccn;tfuc:d~cço¡dir.g!ö!::e

known amir;o :i::id ~~'q:.t:icc of BPTl_ The auihcr, netc

¡edhkcccdlpwJucÜc;iuystiii",¡'uii-gihc(:mwc¡"io1\ vi "rimiiivc pu,curwr cdl$ iii ih~ bo~" ""r,,(,)w ínil'
pi(~¡:-1hmb!~!~ which ~tb'c(;\l"ndy i:l"~\.re, "ynth:.
~¡:e hemo¡;hJ.iín ¡u~d ;lfi: r::k:i~cd into il:c cir;..LI:::íc,r. ::,
ri:bl()üd.;dJ: \\'hen the nDi:btr c:'re:!

" determin"i~()fl of por prc~pect'; for iwbtinf; mRNA
~iiii.~bic for syi\the.,is Df:= cDN,.\ iihr;iry d~,,, 10 .1pP.H"lii

ll'w 1:;\,,,1\ d fi:RNA iii i"iiiall" l::tr.ct\i l';:wiìu gia~IJ ;5
;,:iJ ¡uHf; \h\u,- SOU!(.;: and ihen add!~~ ¡he prNtiecu; of mcce",~ in prohing:' genomic Hbr;;fY i:,iri¡: ~ mixl\r~
of hbelle,j prüh¡:!., 51;iing: '"\loH: generally, mixed.

hlo-:idi:dltin

drcukilIiC,n i~ gre.1ler ili:\ l\ted"d fm t'.0rm~l :i,$'Je,

$.qucntt oi:¡¡ü(\c(lxynucleotitk prabc'Stuvc bttn us:d to íwl;iii: r:¡(¡¡::n ~e¡¡~., ûf unknown ;.:itl"r,c~ frcm .~J
cDN..; ¡¡br~ric~. $'."ch prchc~ :=n: i¡.ric.,li;.~ i,-,i~nlr~~ nr

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