TRNG I HC CN TH

VIN NGHIN CU V PHT TRIN CNG NGH SINH HC ----------------------------------

BI BO CO TM HIU V SNP (SINGLE NUCLEOTIDE POLYMORPHISMS)

CN B GING DY: GS.TS NGUYN TH LANG VIN LA NG BNG SNG CU LONG NHM HC VIN THC HIN: NHM 5 LP: CAO HC CNSH K19

Cn Th, thng 10 nm 2012

SNP (SINGLE NUCLEOTIDE POLYMORPHISMS)
I. GII THIU V SNP - SNPs (c l snip) c vit tt t ch Single Nucleotide Polymorphisms, c ngha l cc dng a hnh ca nucleotide n. Marker ny thng c dng phn tch genome ngi, hin nay c p dng cho genome ca nhiu sinh vt khc vi s lng ln nht t trc n nay, nh mt t bin im ti mt nucleotide trn genome (hnh 1).

Hnh 1: Mt t bin im xy ra dn n thay th mt cp nucleotide G C bng A T hoc ngc li to ra mt SNP. - a hnh n nucleotide, hoc SNPs l nhng bin th trnh t DNA xy ra khi mt n nucleotide (A, T, C, hoc G) trong trnh t b gen b thay i. V d mt SNP c th thay i trnh t DNA GGCTAA A n A GGCTAA T. i vi mt bin th c coi l mt SNP, n phi xy ra trong t nht 1% dn s. SNPs, to nn khong 90% ca tt c cc bin th di truyn ca con ngi, xy ra mi 100 n 300 cn c dc theo h gen ca con ngi 3-t-base. Hai trong s ba mi SNPs lin quan n s thay th ca cytosine (C) vi thymine (T). SNPs c th xy
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ra trong khu vc ca b gen m ha (gen) v khng m ho. Nhiu ngi SNPs khng c nh hng n chc nng t bo, nhng cc nh khoa hc tin rng nhng ngi khc c th predispose ngi bnh hoc nh hng n phn ng ca h vi mt loi thuc. - SNPs din ra bnh thng trong sut DNA ca mt ngi. Trung bnh xy ra mt ln trong mi 300 nucleotide, c ngha l c khong 10 triu SNPs trong h gen ca con ngi. Thng thng nht, nhng bin th ny c tm thy trong DNA gia cc gen. N c xem nh l nh du sinh hc, cc nh khoa hc gip xc nh v tr cc gen lin quan n bnh. Khi SNPs xy ra trong gen hoc trong mt khu vc gn mt gen quy nh, n c th c vai tr trc tip n s xut hin bnh bng cch nh hng n chc nng ca gen. - Mc d hn 99% trnh t ADN ca con ngi u ging nhau, s thay i trong chui DNA c th c tc ng ln n vic lm th no con ngi bnh, cc yu t mi trng, chng hn nh vi khun, virus, c t, cc ha cht, cc loi thuc v cc liu php iu tr khc. iu ny lm cho SNPs c gi tr cho nghin cu y sinh hc v pht trin cc sn phm dc phm hoc chn on y khoa. SNPs cng tin ha n nh, khng thay i nhiu t th h ny sang th h khc lm cho chng ta d dng hn khi nghin cu dn s. - SNP l nhng bin dng ca chui DNA c tm thy vi tn sut cao trong genome ngi (Taillon Miller v ctv. 1998). Chng ta c th s dng SNP marker phn lp cc yu t di truyn c lin quan n tnh trng bnh l v cng phc tp (Taillon Miller v ctv. 1999). Ngi ta c th d on 100.000 hoc nhiu hn na SNP marker (trong qung 30-kb, hoc 5 marker cho mt gen) trong genome ngi (Collins v ctv. 1997). Nhng phng php nh gi kiu gen vi kt qu cao i hi mt kin thc v chui trnh t rt chnh xc ca SNP. Do , bt c cng b no v SNP phi hm cha hai ni dung:
(1) (2)

Xc nh chui trnh t DNA. Tn s alen. y l mt k thut c p dng vo u nhng nm 2000, t genome ngi n genome cc loi sinh vt khc nh cy Arabidopsis, cy la (Oryza sativa L.),
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II. PHNG PHP XC NH SNP - C hai phng php to ra SNP, mt l dng trc tip m trnh t di truyn v th hai l phn bit cc t bin im thng qua dng tch sc k lng (DHPLC). Thng thng dng primer thit k m trnh t v cc on khuyt i khong 500 cp base. Chng ta c th dng phng php PCR tch hai c th v trn cc c th ny chung, sau un nng v lai thnh lp cc dulex tng ng v d bit. Cc bc phn tch axit nucleic bng SNP nh sau: + Phn lp DNA nn + Chn la primer v thit k primer cho khuch i trong PCR + Tch SNP trn bc sng tng thch (DHPLC) + Phi hp s liu, m trnh t DNA v cloning - Theo phn tch chi tit chui trnh t ca nhng phn no trong genome, nhng trnh t DNA ny t hai c th khc nhau phn ln u ging nhau, vi s cp base khc bit nhau nm trong khong cho php 500 1000bp. Mt cp base ti v tr no biu th s khc nhau ca c th c tnh cht rt ph bin v mt cp base khc l variant t ph bin hn cng mt v tr. Nu cp base c tnh cht t ph bin hn xut hin xut hin vi xc sut nh hn 1% trong qun th, ngi ta nh ngha v tr ca cp base l v tr mt SNP. Hin nay, ngi ta cng b 3.000.000 SNP trong genome ngi (Rusell 2002), nhiu hn bt c DNA marker c cng b trc . Kiu a hnh nh vy v cng quan trng trong di truyn ngi v chng i din cho hn 98% tt c a hnh DNA. Cc alen ca mt SNP c th d dng c xem xt bi phn tch lai vi phn t oligonucleotide no . + Trong quy trnh ny, mt on d (probe) ngn hay on oligonucleotide (gi l phn t oligo) cha khong 25bp c tng hp sao cho n s b sung vo alen ph bin ti locus SNP. Phn t DNA ny c trn vi DNA mc tiu v tin trnh lai c hon chnh mc cc k chnh xc (high stringency). iu ny c ngha l mi iu kin y phi thun li cho s lai gia probe v DNA. + Khi hin tng lai xy ra, DNA mc tiu s c alen ph bin (common allene). Di nhng iu kin cc k chnh xc trong s kin matching (cp
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base tng ng bt cp vi nhau, phn t oligo s khng lai vi DNA mc tiu c mt cp base no b thay i mt SNP (hnh 2).

Phn t lai n nh vi tng cp base hon ton tng ng

Oligonucleotide

G C C A T T A A GT C T T C A T C C C T A C GG T A A T T C A G AA GT A GG G A T
DNA mc tiu

SNP
Mt cp base khng tng ng, hydrid khng n nh Mismatch: lm cho cp base khng hnh thnh c

G C C A T T A A GT

TT CAT CCCTA

C GG T A A T T C AC AA GT A GG G A T
Hnh 2: Phn tch lai oligonucleotide to ra mt SNP. Mt oligonucleotide c tnh cht b sung hon ton i vi alen bnh thng, n c lai vi DNA mc tiu di iu kin ti u c tnh cht fullmatch (bt cp base hon ton tng xng) gia probe v DNA. Khi s kin lai xy ra, DNA mc tiu c alen bnh thng, nhng khi s kin lai khng xy ra, DNA mc tiu c base trong tnh trng mismatch, ngi ta gi l mt a hnh ca SNP. - Vic to ra phn ng cng mt lc hng trm SNP c th c thc hin nh s dng DNA microarray, cn c gi l DNA chips, GeneChips arrays (thng hiu ca Affmetrix, Inc) v oligonucleotide arrays. Microarray c kh nng c bit 100.000 hoc nhiu hn na phn t DNA trn mt din tch 3,6cm2 (hnh 3). + DNA microarray l k thut sp xp v phi hp phn t bao gm cc on DNA v protein cho lai vi nhng phn t probe trong th vin DNA c sp xp theo matrix. K thut DNA microarray nhm mc ch tm hiu s th
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hin gen, mc a dng ngun di truyn, bn gen, phn tch bin ng trong gen. + DNA microarray c hiu mt cch n gin l sp xp mt cch th t hng nghn n v phn t DNA probe vi trnh t ging nhau. Chng c c nh trn mt tm knh hoc trn mng nylon.

Hnh 3: Xc nh a hnh nucleotide n bng k thut microarray. * Lch s ra i ca k thut microarray: Vo cui nhng nm 1960, Pardue & Gall; Jones & Roberson tm ra phng php lai in situ c s dng mu d nh du hunh quang, FISH. Phng php c nh cc nhim sc th v nhn trn phin knh (sao cho DNA to thnh mch kp vi mu d) ngy nay c s dng t DNA ln phin knh trong phng php microarray. Vo thi gian ny, ho hc hu c cng pht trin, cho php tng hp t ng cc mu d oligonucleotide vo nm 1979. K thut phn tch s dng phng php gn ng thi nhiu trnh t ch ln mt b lc hay mng theo th t, phng php thm im (dot blot), c Kafatos v cng s (1979) a ra. Vi k thut ny, cc trnh t ch c c nh trn vt v lai vi mu d (thng l trnh t axit nucleic nh du). Saiki v
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cng s (1989) a ra mt cch khc, dot blot ngc, trong gn nhiu mu d theo th t trn mng v ch phn tch c nh du. Cng thi gian ny, cc array u tin vi gi th khng thm nc c to ra trong phng th nghim ca Maskos (1991). u nhng nm 1990, k thut nh du pht hunh quang a mu c Ried v cng s; Balding & Ward gii thiu. Vo nm 1993, array cha cc oligonucleotide ngn, di 19 nucleotide c tng hp in situ. Nm 1994, Hoheisel v cng s tng mt chm (spot) bng cch dng robot ly v t mu d ln gi th. Phng php t ng ho ny lm tng tc qu trnh, gim cc sai st chc chn mc phi khi thc hin nhng th tc c tnh lp li cao bng tay, v tng tnh chnh xc v tr, tng tnh ng hnh ca cc spot mu. Tt c cc th nghim tin phong trn l c s ca k thut array hin nay. Ngi ta nh gi rng, k thut ny c th s pht trin n mc ch vi nm na c th so snh n vi k thut PCR khng th thiu trong sinh hc hin nay. * Trnh t thc hin microarray c m t nh sau: (1) Thc hin cng vic in cc slide, chy PCR, thit lp chng trnh m bo cht lng sn phm PCR, lm tinh khit sn phm PCR. (2) Sao chp microarray thng qua h mt thng t ng, to ra h s chi tit v chun b gen. (3) Thc hin lai DNA vi probe, trong ngi ta cn phi ly trch RNA nhum vi Cy3 v Cy5. (4) Thc hin vic chp hnh thng qua my scanner v phn tch s liu. Th nghim trong DNA microarray cn c nhng probe v DNA mc tiu, trong , probe l nhng phn t DNA c nh trn chip bng thy tinh hoc silicon, chng cha c nh du, nhng DNA mc tiu c nh du m nng ca n s c phn tch. V vy, ngi ta s dng mt chip tn gi l probe array. Hunh quang c s dng nh du l florescein hoc cyanine, l phm mu Cy3 c mu xanh l cy v phm mu Cy5 c mu . Du hiu nh du bng microarray chng ta phi dng hai cht mu nhum ph bin l Cy3 v Cy5. Cc cht nhum ny thng l dUTP hoc dCTP. Cy3
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cho mu vng cam vi nh sng hp thu ti thiu 50nm v ti a 581nm. Cy5 cho mu vi nh sng hp thu ti thiu l 649nm v ti a l 670nm. Kt qu hnh nh s th hin cc gen mc tiu c nhn bit trong matrix. Nu m microarray biu hin c gen mc tiu, kt qu dng tnh. Ngc li, phn ng s m tnh (khng c kt qu lai vi probe).

Hnh 4: Quy trnh ca microarray. Nhiu k thut c nghin cu to ra SNP theo kiu t ng ha vi s gip ca computer (Wang v ctv. 1998). Nhng k thut l: EST Sequence Data, EST Assembly vi phn mm chuyn trch, phng php nh gi kiu gen thng qua thit k mi (primer) cho PCR v GBA theo phn mm GBAprimer1.1, phng php xc nh SNP bng k thut c chui trnh t pyrophosphate DNA vo thi im thch hp (Alderborn v ctv. 2000) III. TNH CHT CA SNP - SNP c tnh cht diallelic trong qun th v tn s alen ca n c th c c on d dng trong bt c qun th no, thng qua mt lot xt nghim k thut (Kwor v ctv. 1994). - SNP l nhng marker c tnh n nh rt cao v mt di truyn.
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- L sn phm ca PCR. - c tm thy vi tn sut cao nht trong genome ngi. - a dng cao. IV. NG DNG V TRIN VNG CA SNPs 1. SNPs bn - Ngi ta pht hin mt chin lc nghin cu gip cho vic pht hin nhanh chng nhng SNP t s liu lu tr EST (Expressed Sequence Tag) (Picuolt Newberg v ctv. 1999). S kin pht trin in vitro nhm khuch i nhng trnh t v tr c bit, v d nh PCR v khm ph marker c tnh a hnh v c thng tin di truyn cao nh microsatallite, STR (short tandem repeat), s kin nh vy v ang to iu kin thun li chng ta sng to ra nhng bn di truyn c mt thp (lom density maps) ca ngi, ng dng c hiu qu trong lnh vc y khoa, th d xt nghim bnh u x, bnh Huntington, bnh tiu ng, (Broman v ctv. 1998). - Cc nh khoa hc tin rng SNP bn s gip h xc nh c nhiu gen lin quan vi cc bnh phc tp nh ung th, bnh tiu ng, bnh mch mu, v mt s hnh thc ca bnh tm thn. - Mt vi nhm lm vic tm SNPs v cui cng to ra SNP bn h gen ca con ngi. Trong s l M Human Genome Project (HGP) v mt nhm ln ca cc cng ty dc phm c gi l SNP Consortium, d n TSC. Kh nng trng lp gia cc nhm nh l kh xy ra v c khong 3 triu SNPs, v phn thng tim nng ca mt bn SNP l cao. - Ngoi ngha nghin cu pharmacogenomic trong chn on, y sinh hc, SNP bn cn gip xc nh hng ngn cc du hiu b sung trong b gen, do hng nghin cu ca bn b gen l rt ln. 2. Lm th no SNPs c th c s dng nh l yu t nguy c pht trin bnh? - SNPs khng gy bnh, nhng n c th gip xc nh kh nng rng ai s pht trin mt cn bnh c th. Mt trong nhng gen lin quan vi bnh Alzheimer, apolipoprotein E hoc APOE, l mt v d tt v cch SNPs nh hng n s pht trin ca bnh apoE cha hai SNPs m kt qu trong ba alen c th
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cho gen ny: E2, E3, E4. Mi allele khc nhau bi mt c s DNA, v cc sn phm protein ca mi gen s khc bi mt amino axit. - Mi c nhn c tha hng mt bn sao m ca APOE v mt bn sao ni ca APOE. Nghin cu cho thy rng mt ngi c tha hng t nht mt E4 allele s c mt c hi ln pht trin bnh Alzheimer. R rng, s thay i ca mt amino acid trong cc protein E4 lm thay i cu trc v chc nng ca protein v c nhiu kh nng lm cho bnh pht trin. Nu mt ngi k tha allele E2 th t c kh nng pht trin bnh Alzheimer. - Tt nhin, SNPs khng phi l ch s tuyt i ca s pht trin ca bnh. Mt ngi no c tha hng hai alen E4 khng bao gi c th pht trin bnh Alzheimer, trong khi mt ngi c tha hng hai alen E2 c th. APOE ch l mt gen c lin quan n bnh Alzheimer. Ging nh cc ri lon mn tnh nh bnh tim, tiu ng, hoc ung th ph bin nht, bnh Alzheimer l mt bnh c th c gy ra bi cc bin th trong mt vi gen. - Hu ht cc SNPs khng c nh hng n sc khe hoc s pht trin. Mt s nhng khc bit di truyn c chng minh l rt quan trng trong vic nghin cu v sc khe con ngi. Cc nh nghin cu tm thy SNPs c th gip d on phn ng ca mt c nhn vi mt s loi thuc nht nh, nhy cm vi cc yu t mi trng nh cht c, v nguy c pht trin cc bnh c th. SNPs cng c th c s dng theo di cc tha k ca cc gen bnh trong gia nh. Nghin cu trong tng lai s lm vic xc nh SNPs lin kt vi cc bnh phc tp nh bnh tim, tiu ng v ung th. - Nhiu loi bnh ph bin ngi khng c gy ra bi mt bin th di truyn trong mt gen duy nht, nhng b nh hng bi cc tng tc phc tp gia nhiu gen cng nh cc yu t mi trng v li sng. - Mc d cc yu t mi trng v li sng c nh hng n s pht trin mt cn bnh, y l kh khn o lng v nh gi hiu qu tng th ca h v qu trnh bnh. V vy, tim nng ca mt c nhn pht trin mt bnh da trn gen v cc yu t di truyn. - Yu t di truyn cng c th trao nhy cm hoc khng bnh v xc nh mc nghim trng hoc tin trin ca bnh. Bi v chng ta vn cha bit c
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tt c cc yu t lin quan trong nhng ng phc tp, cc nh nghin cu tm thy n kh khn pht trin cc xt nghim sng lc cho hu ht cc bnh v cc ri lon. Bng cch nghin cu mi lin quan ca SNP vi mt c im bnh, cc nh nghin cu pht hin cc gen c lin quan vi mt cn bnh. Xc nh v hiu bit v vai tr ca yu t di truyn trong bnh cng s cho php cc nh nghin cu nh gi tt hn vai tr ca yu t di truyn, chng hn nh hnh vi, ch n ung, li sng, v hot ng th cht c trn bnh. - Bi v yu t di truyn cng nh hng n phn ng ca mt ngi iu tr bng thuc, a hnh DNA chng hn nh SNPs s l hu ch trong vic gip cc nh nghin cu xc nh v hiu l do ti sao cc c nhn khc nhau trong kh nng ca mnh c th hp th cc loi thuc nht nh, cng nh xc nh l do ti sao mt c nhn c th tri nghim mt tc dng ph bt li cho mt loi thuc c th. V vy, vic pht hin gn y ca SNPs ha hn mang n mt cuc cch mng khng ch l qu trnh pht hin bnh nhng thc t thuc phng nga v cha bnh. 3. SNPs v chn on bnh: - Vt liu di truyn ca mi ngi c mt m hnh SNP c o c to thnh nhiu bin th di truyn khc nhau. Cc nh nghin cu tm thy rng hu ht cc SNPs khng chu trch nhim cho mt bnh tt no. Thay vo , n nh l cc du hiu sinh hc xc nh chnh xc mt cn bnh trn bn h gen ca con ngi, bi v n thng nm gn mt gen c tm thy c lin quan n mt cn bnh no . Thnh thong, mt SNP thc s c th gy ra mt cn bnh, v do , c th c s dng tm kim v c lp cc gen gy bnh. to mt th nghim di truyn, gen gy bnh no c xc nh bi cc nh khoa hc thu thp mu mu t mt nhm cc c nhn b nh hng bi cn bnh ny v phn tch DNA ca h cho cc mu SNP. Tip theo, cc nh nghin cu so snh cc m hnh vi cc mu thu c bng cch phn tch ADN t mt nhm cc c nhn khng b nh hng bi cn bnh ny. Loi so snh ny, c gi l "Hip hi nghin cu", c th pht hin s khc bit gia cc m hnh SNP ca hai nhm, qua cho thy l m hnh rt c th lin quan vi gen gy bnh. Cui cng, h s SNP l c trng ca nhiu loi bnh khc s c
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thnh lp. Sau , ch l vn thi gian, cc bc s c th xc nh mt ngi c th nhy cm vi mt cn bnh no ch bng cch phn tch cc mu DNA ca h cho m hnh c th SNP. 4. Pht trin SNPs v dc: - Nh cp trc , SNPs cng c th c kt hp vi hp th v gii phng mt bng ca cc i l tr liu. Hin nay, khng c cch no n gin xc nh lm th no mt bnh nhn s p ng vi mt loi thuc c th. Mt iu tr c chng minh c hiu qu trong mt bnh nhn nhng c th khng c hiu qu nhng ngi khc. T hn na, mt s bnh nhn c th tri qua mt phn ng min dch bt li ca mt loi thuc c th. - Trong tng lai, cc loi thuc thch hp nht cho mt c nhn c th c xc nh trc iu tr bng cch phn tch h s SNP ca bnh nhn. Mc tiu l to mt loi thuc c kh nng ph hp cho c nhn , c gi l "thuc c nhn ho", iu ny s cho php cc cng ty dc phm mang li nhiu loi thuc hn cho th trng v cho php cc bc s k toa cc phng php iu tr cho tng c nhn c th theo nhu cu ca bnh nhn. 5. SNPs v NCBI Bi v SNPs xy ra thng xuyn trong b gen v c xu hng tng i n nh v mt di truyn, n l ch du sinh hc tuyt vi. nh du sinh hc l cc on DNA vi mt v tr a l, nhn dng c th d dng theo di v c s dng xy dng bn nhim sc th cho thy v tr ca cc gen bit c, hoc cc du hiu khc, tng i vi nhau. Nhng bn ny cho php cc nh nghin cu nghin cu v xc nh nhng c im kt qu t s tng tc ca nhiu gen. NCBI ng mt vai tr quan trng trong vic to iu kin thun li cho vic xc nh v danh mc SNPs thng qua vic to ra v duy tr c s d liu SNP cng cng ( dbSNP ). Cng c mnh m di truyn c th c truy cp bi cc cng ng y sinh hc trn ton th gii v c d nh kch thch nhiu lnh vc nghin cu sinh hc, bao gm c vic xc nh cc thnh phn di truyn ca bnh. NCBI "Khm ph khng gian" To iu kin thun li cho SNP nghin cu. Cc bn ghi trong dbSNP cross-ch thch trong cc ngun thng tin ni b khc
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nh PubMed, trnh t d n b gen, GenBank h s, c s d liu Gene Entrez, v c s d liu dbSTS. Ngi dng c th truy vn dbSNP trc tip hoc bt u mt tm kim trong bt k phn no ca khng gian NCBI khm ph xy dng mt b h s dbSNP p ng cc iu kin tm kim ca h. H s cng c tch hp vi cc ngun thng tin bn ngoi thng qua siu vn bn URL m ngi dng dbSNP th lm theo tm hiu thng tin chi tit l vt ra ngoi phm vi ca curation dbSNP. to iu kin thun li cho cc n lc nghin cu, NCBI ca dbSNP c bao gm trong h thng phc hi Entrez m cung cp truy cp tch hp mt s cng c phn mm v c s d liu c th h tr trong phn tch SNP. V d, mi bn ghi SNP trong cc lin kt c s d liu cc ngun lc b sung trong "V tr Discovery ca NCBI. Ti nguyn bao gm: GenBank , c s d liu trnh t NIH ca Gene Entrez im mt u mi cho cc gen v cc thng tin lin quan; dbSTS , NCBI ca ngun ti nguyn c cha chui v d liu bn trn mc ngn gen; b gen ca con ngi sp xp d liu; v PubMed , tm kim ti liu NCBI v h thng phc hi. SNP h s cng lin kt n cc ngun lc bn ngoi khc nhau c lin quan. Cung cp truy cp n mt trang web "mt ca SNP mua sm" to iu kin nghin cu khoa hc trong nhiu lnh vc khc nhau, t di truyn qun th v sinh hc tin ha quy m ln bnh v nghin cu n thuc. u t lu di trong cc nghin cu ha hn khng ch nng cao sinh hc ca con ngi m cn cch mng ha trong thc hnh y hc hin i. TI LIU THAM KHO 1. PGS.TS Nguyn Th Lang v GS. TS Bi Ch Bu. 2005. Sinh hc phn t gii thiu phng php v ng dng. Nh xut bn Nng Nghip & Pht trin Nng thn. Trang 72-73. 2. GS. TS Bi Ch Bu v PGS.TS Nguyn Th Lang. 2008. Di truyn phn t. 2008. Nh xut bn Nng Nghip & Pht trin Nng thn. Trang 180-184. 3. NCBI c s d liu ca cc a hnh n nucleotide (dbSNP)
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4. C s d liu ca Nht Bn a hnh n nucleotide (JSNP)News Network Genome 5. dbSNP c s d liu - T Trung tm Quc gia v Thng tin Cng ngh sinh hc (NCBI). 6. SNPs: Cc bin th trn mt ch gii thiu c bn SNPs t Trung tm Quc gia v Thng tin Cng ngh sinh hc (NCBI). 7. Hiu SNPs v ung th - Mt hng dn trc tuyn t Vin Ung th Quc gia. 8. Diseases and Medical Response - An animated tutorial describing how DNA markers are used in medical applications. Part of the Kids Genetics website from GlaxoSmithKline. 9. Genome Variations - Questions and answers about genome variation from the Genome News Network. 10. The SNP Consortium Website: Past, Present, and Future 2003. Nucleic Acids Research 31 (1), 124-27. 11. Introduction to SNPs: Discovery of Markers for Disease 2002. BioTechniques (pdf). 12. Prospecting for Gold in Genome Gulch 2002. The Scientist . 13. Single Nucleotide Polymorphisms: From the Evolutionary Past 2001. Nature . 14. Single Nucleotide Polymorphisms: . . .To a Future of Genetic Medicine 2001. Nature .

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