Ky Thuat Dien Di

8/2/2019 Ky Thuat Dien Di

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Ti liu chia s ti: wWw.SinhHoc.edu.vn

I. Khi nimI. Khi nim

II. ng dng ca hin tng in di.II. ng dng ca hin tng in di.

1. ng dng k thut in di protein SDS-PAGE ci thin1. ng dng k thut in di protein SDS-PAGE ci thinv nng cao phm cht cc ging la.v nng cao phm cht cc ging la.

2.2. in di phn tch ADN v ARNin di phn tch ADN v ARN..

3. in di thuc.3. in di thuc.

4. Phn tch protein bng in di trn gel.4. Phn tch protein bng in di trn gel.

5. in di Protein v DNA trn cy bi Da xanh.5. in di Protein v DNA trn cy bi Da xanh.

6. ng dng k thut in di Isozyme trong cng tc nh6. ng dng k thut in di Isozyme trong cng tc nh

danh ging cao su vn nhn tc ghp.danh ging cao su vn nhn tc ghp.7. Dng phng php in di i vi da7. Dng phng php in di i vi da

8. My in di8. My in di


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I. Khi nimI. Khi nim

in di: S di chuyn ca cc ht mang in diin di: S di chuyn ca cc ht mang in ditc dng ca in trng v chiu chuyn ngtc dng ca in trng v chiu chuyn ngca n v in cc tri du.ca n v in cc tri du.

in di hayin di hay in di trn gelin di trn gel ((electrophoresiselectrophoresis hayhaygel electrophoresisgel electrophoresis) p dng trong sinh hc phn t) p dng trong sinh hc phn tl mt k thut phn tch cc phn tl mt k thut phn tch cc phn tADNADN,, ARNARNhay proteihay proteinn da trn cc c im vt l ca chngda trn cc c im vt l ca chngnh kch thc, hnh dng hay im ngnh kch thc, hnh dng hay im ng in tin tchch(isoelectric point).(isoelectric point).


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II. ng dng ca hin tng in di.II. ng dng ca hin tng in di.

1.1. ng dng k thut in di protein SDS-ng dng k thut in di protein SDS-PAGE ci thin v nng cao phm chtPAGE ci thin v nng cao phm chtcc ging la.cc ging la.


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in di protein SDS-PAGE (Sodiumin di protein SDS-PAGE (Sodium

Dodecyl Sulfate PolyAcrylamide) c th ngDodecyl Sulfate PolyAcrylamide) c th ngdng trn thc vt, ng vt, vi sinh vt.dng trn thc vt, ng vt, vi sinh vt.

Trong chn to ging la, k thut nyTrong chn to ging la, k thut nygip pht hin nhanh nhng tnh cht ni btgip pht hin nhanh nhng tnh cht ni btnh mi thm, protein, amylose...nh mi thm, protein, amylose...


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Bung in di protein


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Phn tch protein bng in di SDS-PAGE


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Ti liu chia s ti: wWw.SinhHoc.edu.vn Ci thin phm chtCi thin phm chtnhng ging c snnhng ging c snnhnh

Np B 1-2Np B 1-2 c chtc chtlng cm nu ngon,lng cm nu ngon,do, chiu di ht vdo, chiu di ht vnng sut cao hnnng sut cao hn

ging Np B trcging Np B trcy nh ng dng ky nh ng dng kthut in di proteinthut in di proteinSDS-PAGE phcSDS-PAGE phctrng ging la Np Btrng ging la Np B

c sn ti tnh Tinc sn ti tnh TinGiang.Giang. Np B 1-2


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Ti liu chia s ti: wWw.SinhHoc.edu.vnJasmine 85Jasmine 85 c biu hin bc bng, mi thmc biu hin bc bng, mi thmkhng n nh, kh tm c th trng tiukhng n nh, kh tm c th trng tiu

th.th.Nhng sau khi ging Jasmine c ci thinNhng sau khi ging Jasmine c ci thin

phm cht bng k thut in di protein SDS-phm cht bng k thut in di protein SDS-

PAGE, cc ging ny pht trin tt, t su bnh,PAGE, cc ging ny pht trin tt, t su bnh,c th t nng sut v cht lng cao.c th t nng sut v cht lng cao.


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Ti liu chia s ti: wWw.SinhHoc.edu.vnK thut in di protein SDS-PAGE gipK thut in di protein SDS-PAGE gipthanh lc cc dng b thoi ha v tuynthanh lc cc dng b thoi ha v tuynchn ging theo hng ci thin phmchn ging theo hng ci thin phmcht cm nu, lm mm cm. c bit lcht cm nu, lm mm cm. c bit ltng hm lng protein, gip ngon cmtng hm lng protein, gip ngon cmhn. K thut ny c chi ph thp, cho kthn. K thut ny c chi ph thp, cho kt

qu nhanh hn so vi phng php chnqu nhanh hn so vi phng php chnlc truyn thng. Kh nng chng chulc truyn thng. Kh nng chng chusu bnh hi ca cc ging ny tng isu bnh hi ca cc ging ny tng itt. Trong cc v qua, c mt s t rytt. Trong cc v qua, c mt s t ry

nu di tr nhng la khng b nh hngnu di tr nhng la khng b nh hngnhiu. Cc ging ang tr bng u nnnhiu. Cc ging ang tr bng u nnnng sut cao.nng sut cao.


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Vic to ra ging la thch nghi vi iuVic to ra ging la thch nghi vi iukin mn xm nhp vo t lin bng kkin mn xm nhp vo t lin bng k

thut in di SDS-PAGE protein phtthut in di SDS-PAGE protein phthin c cc ging ch th thm v chuhin c cc ging ch th thm v chumn c th ng dng ngay cho cng tcmn c th ng dng ngay cho cng tclai chn.lai chn.


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K thut in di protein SDS-PAGE


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Ti liu chia s ti: wWw.SinhHoc.edu.vn2.2.in di phn tch ADN v ARNin di phn tch ADN v ARN

Nguyn tc ca phng php l do: di tc ngNguyn tc ca phng php l do: di tc ng

ca in trng, cc phn t ADN (thng tchca in trng, cc phn t ADN (thng tchin m) khc nhau v kch thc, in tch, mcin m) khc nhau v kch thc, in tch, mc cun xon v dng phn t (mch thng hay cun xon v dng phn t (mch thng haymch vng) s di chuyn qua h mng ca gel tmch vng) s di chuyn qua h mng ca gel t

cc m (cathode) sang cc dng (anode) vi tccc m (cathode) sang cc dng (anode) vi tc di chuyn khc nhau. V vy, di chuyn khc nhau. V vy, chng dn dnchng dn dntch nhau ra trn trng in di, qua ngi ta ctch nhau ra trn trng in di, qua ngi ta cth thu thp v phn tch c tng phn onth thu thp v phn tch c tng phn on

ADN hoc gen ring r. Trn in trng cc phnADN hoc gen ring r. Trn in trng cc phnon ADN c kch thc cng nh cng c tc on ADN c kch thc cng nh cng c tc di chuyn trn in trng nhanh hn.di chuyn trn in trng nhanh hn.


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Sau khi in di kt thc, cc phn t ADN cSau khi in di kt thc, cc phn t ADN cth quan st thy nh s dng cc thucth quan st thy nh s dng cc thucnhum pht hunh quang, chng hn nhnhum pht hunh quang, chng hn nh

ethidium (cht ny gn kt vi ADN bngethidium (cht ny gn kt vi ADN bngcch ci vo khe gia cc nucleotit). Micch ci vo khe gia cc nucleotit). Mimt bng in di thng phn nh mt tpmt bng in di thng phn nh mt tphp cc phn t ADN c cng kch thc.hp cc phn t ADN c cng kch thc.


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in di gel agarose cc ch phm ADN thc vtA: Trung Qun , B: Qu B,C: Lc Vng, D: Sng My.

M: Thang chun ADN l HindIIIM: M , CT: Ct Tin


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in di gel agarose cc sn phm RAPD-PCR ca mu

Trung Qun vi cc mi D5, D6, D7, OPA4, OPA10 vOPA12M: Thang chun ADN 1 kb, M: M , CT: Ct Tin


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in di gel agarose sn phm RAPD-PCR ca mu Qu B vi ccmi OPA4, OPA10 v OPA12M: Thang chun ADN 1 kb, M: M , CT: Ct Tin


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in di gel agarose sn phm RAPD-PCR ca mu Lc Vng vi ccmi P4, P10, Z16, Z17, Z18 v Z19.M: Thang chun ADN 1 kb, M: M , CT: Ct Tin


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in di gel agarose sn phm RAPD-PCR ca mu SngMy vi cc mi P4, P10, Z16, Z17, Z18 v Z19.M: Thang chun ADN 1 kb, M: M , CT: Ct Tin


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in di trn gel agaroza 1% kim tra ccplasmit ti t hp


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Ti liu chia s ti: wWw.SinhHoc.edu.vn** ng dng:ng dng:in di huyt sc t gip cho vic xcin di huyt sc t gip cho vic xcnh cc huyt sc t khng bnh thng gy ranh cc huyt sc t khng bnh thng gy ra

nhiu bnh l...), bnh nh: bnh ri lon huyt scnhiu bnh l...), bnh nh: bnh ri lon huyt sct v s lng (beta- thalassemia, denta-thalassemiat v s lng (beta- thalassemia, denta-thalassemiari lon huyt sc t v cht lng (bnh huyt scri lon huyt sc t v cht lng (bnh huyt sct S, M, E,...). y l nhng bnh thiu mu huytt S, M, E,...). y l nhng bnh thiu mu huyt

tn bm sinh bt thng v huyt sc t .tn bm sinh bt thng v huyt sc t .


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in di xc nh huyt thngin di xc nh huyt thng

i i i i Si


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IN DI RA CC VCH DI NGN KHC NHAU. HNH

MINH HA CA CC ON DNA

Ti li hi i W Si hH d


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Ti liu chia s ti: wWw.SinhHoc.edu.vn3.3. in di thucin di thuc

Khi nimKhi nim..

in di thuc (electrophoresis) l phng php dngin di thuc (electrophoresis) l phng php dngdng in mt chiu u y mt s ion thucdng in mt chiu u y mt s ion thuciu tr vo c th hoc ly cc ion thuc c hi raiu tr vo c th hoc ly cc ion thuc c hi rakhi c th.khi c th.

** Nguyn l ca phng php in diNguyn l ca phng php in di thucthuc l: Ccl: Ccin tch cng du th y nhau, tri du th ht nhau.in tch cng du th y nhau, tri du th ht nhau.Mt dung dch m trong cht ha tan c th phnMt dung dch m trong cht ha tan c th phnly thnh cc ion (gi l dung dch in ly), khi cly thnh cc ion (gi l dung dch in ly), khi ca vo mt in cc v cho dng in mt chiua vo mt in cc v cho dng in mt chiuchy qua th cc ion cng du vi in cc s bchy qua th cc ion cng du vi in cc s by ra khi in cc i vo c th.y ra khi in cc i vo c th.

Ti li hi t i W Si hH dN l i l i h i ( N l i l t i h i (


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Ti liu chia s ti: wWw.SinhHoc.edu.vn Ngc li nu mun ly mt ion c hi (vNgc li nu mun ly mt ion c hi (vd ion Ca+2) ra khi c th th ta t ind ion Ca+2) ra khi c th th ta t incc tri du vo vng da nhim ion, incc tri du vo vng da nhim ion, in

cc s ht cc ion ny ra khi c th vcc s ht cc ion ny ra khi c th vpha n.pha n.

Ti li hi t i W Si hH d


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Ti liu chia s ti: wWw.SinhHoc.edu.vn-- Tc dng ca in di thuc gm tc dng doTc dng ca in di thuc gm tc dng do

dng in mt chiu u v tc dng do iondng in mt chiu u v tc dng do ion

thuc gy ra.thuc gy ra.Tc dng ca dng in 1 chiuTc dng ca dng in 1 chiu uu

++ Tc dng gin mch: Ti vng da t inTc dng gin mch: Ti vng da t in

cc c vi m s c hin tng da docc c vi m s c hin tng da dogin mch v c th ko di hng gi. Tcgin mch v c th ko di hng gi. Tcdng gin mch s lm tng cng tundng gin mch s lm tng cng tun

hon v dinh dng, tng chuyn ha,hon v dinh dng, tng chuyn ha,chng vim. C ch do dng in tc dngchng vim. C ch do dng in tc dngtrc tip ln h thn kinh vn mch.trc tip ln h thn kinh vn mch.



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++ Tc dng ln h thn kinh: Ti cc dngTc dng ln h thn kinh: Ti cc dngc tc dng gim au, gim co tht, gimc tc dng gim au, gim co tht, gimtrng lc c. Ti cc m: c tc dngtrng lc c. Ti cc m: c tc dng

kch thch, lm tng trng lc ckch thch, lm tng trng lc c..

Tc dng ca ion thucTc dng ca ion thuc

Ti li hi t i W Si hH dDung dch thucDung dch thuc Ion tc dngIon tc dng CcCc Tc dng iuTc dng iu


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Ti liu chia s ti: wWw.SinhHoc.edu.vnDung dch thucDung dch thuc Ion tc dngIon tc dng CcCc Tc dng iuTc dng iutrtr

Kali-Natri iodure (INaIK)Kali-Natri iodure (INaIK)5%5%

I-I- Lm mm tLm mm tchc sochc so

Natri clorua (NaCl) 5%Natri clorua (NaCl) 5% Cl-Cl- To phn xTo phn xchng vim khpchng vim khp

Kali-Natri bromua (BrNa-Kali-Natri bromua (BrNa-BrK)BrK)

Br-Br- An thnAn thn

Natri salicylat 5%Natri salicylat 5% Salicylat-Salicylat- Chng vimChng vimgim augim au

Novocain 5%Novocain 5% Novocain+Novocain+ ++ Gim auGim au

Hydrocortison acetatHydrocortison acetat Hydrocortison+Hydrocortison+ ++ Chng vimChng vim

NivalinNivalin Nivalin+Nivalin+ ++ Tng cng dnTng cng dn

truyn thn kinhtruyn thn kinh



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Ti liu chia s ti: wWw.SinhHoc.edu.vnCh nh iu trCh nh iu tr

-- Gim au hoc ct cn au trong cc chng auGim au hoc ct cn au trong cc chng au

do nguyn nhn thn kinh, chn thng, co mchdo nguyn nhn thn kinh, chn thng, co mch(chng co tht mch ngoi vi, vim tc ng(chng co tht mch ngoi vi, vim tc ngmch, vim tc tnh mch).mch, vim tc tnh mch).

-- Chng vim mn nh vim khp, vim dy thnChng vim mn nh vim khp, vim dy thn

kinh, vim da, vim mng tip hpkinh, vim da, vim mng tip hp-- Tng dinh dng v tun hon ti ch iu tr vtTng dinh dng v tun hon ti ch iu tr vt

thng vt lot lu lithng vt lot lu linn, so x cng, teo c., so x cng, teo c.

-- To phn x t on iu ha cc ri lon caTo phn x t on iu ha cc ri lon cacc c quan ni tng su v xa ch t incc c quan ni tng su v xa ch t incc. iu ha qu trnh hng phn v c ch cacc. iu ha qu trnh hng phn v c ch cav no.v no.



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4.4. Phn tch protein bng in di trn gelPhn tch protein bng in di trn gel

S tinh sch ca protein rt quan trng v tS tinh sch ca protein rt quan trng v t

protein tinh sch chng ta c th xc nhprotein tinh sch chng ta c th xc nhc trnh t acid amin, mi lin h v tinc trnh t acid amin, mi lin h v tinha gia cc protein trong nhng c th khcha gia cc protein trong nhng c th khc

nhau v kho st cc chc nng sinh hanhau v kho st cc chc nng sinh haca cc protein .ca cc protein .

Ti liu chia s ti: wWw SinhHoc edu vn


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minh ha sc

k trao i ion



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Minh ha c ch gia proteinMinh ha c ch gia proteintrong h sc k trao i iontrong h sc k trao i ion

(a) Nhng ht mang in tch(a) Nhng ht mang in tchdng s trao i ion m vidng s trao i ion m vidung dch m. Protein tchdung dch m. Protein tch

in m cng ion dngin m cng ion dngtng tc vi ntng tc vi n

(b) Khi protein gn vi ht,(b) Khi protein gn vi ht,protein thay th nhng ion mprotein thay th nhng ion m

tng tc vi ht cng nhtng tc vi ht cng nhht thay th nhng ion dnght thay th nhng ion dngtng tc vi proteintng tc vi protein



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Ti liu chia s ti: wWw.SinhHoc.edu.vnMinh ha cho c ch tinh sch Concanavalin AMinh ha cho c ch tinh sch Concanavalin Abng sc k i lcbng sc k i lc

Ti liu chia s ti: wWw SinhHoc edu vn5 in di Protein v DNA trn cy bi Da xanh5 in di Protein v DNA trn cy bi Da xanh


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Ti liu chia s ti: wWw.SinhHoc.edu.vn5. in di Protein v DNA trn cy bi Da xanh5. in di Protein v DNA trn cy bi Da xanhng dng k thut bng phng php phn tchng dng k thut bng phng php phn tch

in di Protein v DNA trn cy bi Da xanh in di Protein v DNA trn cy bi Da xanh xc nh mc a dng di truyn ca ging bixc nh mc a dng di truyn ca ging biDa xanh trong tnh Bn Tre.Da xanh trong tnh Bn Tre.Thit b phn tch nh b ngun chy in di proteinThit b phn tch nh b ngun chy in di protein

SDS-PAGE, k thut DNA v cc ha cht nhumSDS-PAGE, k thut DNA v cc ha cht nhummu,mu,



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6.6.ng dng k thut in di Isozyme trongng dng k thut in di Isozyme trong

cng tc nh danh ging cao su vncng tc nh danh ging cao su vnnhn tc ghpnhn tc ghp

Phng php nh danh truyn thng cc dngPhng php nh danh truyn thng cc dng

v tnh cao su trong vn nhn da vo sv tnh cao su trong vn nhn da vo sphn bit trn 20 ch tiu hnh thi ca tcphn bit trn 20 ch tiu hnh thi ca tcghp, l phng php nh tnh, n gin, choghp, l phng php nh tnh, n gin, chon nay vn c p dng ph bin vi mc tinn nay vn c p dng ph bin vi mc tin

cy kh cao.cy kh cao.



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7. Dng phng php in di i vi da7. Dng phng php in di i vi da-- Siu m bng ion in di dn xut tinh cht voSiu m bng ion in di dn xut tinh cht vo

su trong da, gip da nga v tr mnsu trong da, gip da nga v tr mn



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My lun nhit - thit b tch DNA v RNAtrong k thut PCR ti Bnh vin Da liu

TPHCM



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Ti liu chia s ti: wWw.SinhHoc.edu.vn8. My in di8. My in diMt s loi my in di thng dngMt s loi my in di thng dng

B in di nmngang c trung

H thng chp nhv phn tch gel indi



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H thng FASTReal-time PCR 6

knh

H thng PCRGradient FAST Dual

block 48/48

My bin npxung in



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Ti liu chia s ti: wWw.SinhHoc.edu.vnin di ion dng trong iu tr ra m hi tayin di ion dng trong iu tr ra m hi tay

v chn t tiu chun Chu uv chn t tiu chun Chu u



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