Práctica de Laboratorio 5. Extracción de ADN de Aleta de Pescado

 

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Wråftbfe Mº 1 Jebshm Obkua` Ia `e Fruz Oetah, ^hse Oerïe Ia `e ^hse \e`iaz, ^hiaxy Astcar Fuaves Mhves, A`beme Ob`ame Yajaie Nrbhsh, Lremfcasfe Nåaz Eieoas. @enhrethrbh ia Nbhquïobfe, ^afbmth ^afbmth Lâ`bx Averbst Averbsth h Oajïe BVLHIHVQ (73/5>/>7>7)

^AVQOAM

Hnjatbvh kamare` y aspafïlbfh 5. Ebs`e Ebs`err a` EIM EIM ia ia `e `es s e`ates e`ates ia ia pasfei pasfeih. h.   >. Hn Hnta tama marr `e lhr lhroe oe purbl purblbf bfei eie e ia EIM EIM qua puaia usersa eibfbhme`oamta pere emå`bsbs oh`afu`er. 6. Fh Fhop opra rami miar ar a` prbm prbmfb fbpb pbh h y a` prhf prhfas ash h ia axtreffböm ia EIM.

 

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BMY^HIQFFBÖM A` åf åfbi bih h ia iash shxb xbrr rrbn bnhmu hmuf` f`ab abfh fh (E (EIM IM)) as um åf åfbi bih h muf` muf`ab abfh fh fh fhop op`a `ajh jh qu quaa fh fhmt mtba bama ma a` föibkh kamâtbfh fhm `es bmstruffbhmas pere a` ia iase serr rrh` h``h `h y lumfb lumfbhm hmeo eoba bamt mth h ia thih thihss `hs `hs hrkembsohs vbvhs fhmhfbihs, fhm `e axfapfböm

oaibfbme oaibfb me lhr lhrams amsa. a. Whr aja ajaop` op`h, h, `hs fbamtï fbamtïlbf lbfhs hs bmtrhiufam EIM am fâ`u`es ia emboe`as h p`emtes pere emå`bsbs ia VMW, emå`bsbs ia oatb`efböm ia  EIM, emå`bsbs ia verbefböm ia` múoarh ia fhpbes (FM\) h cbnrbiefböm kamöobfe fhoperetbve (FKC), Vhut Vhutcar carm m N`ht N`httb tbmk mk,, WF WF^, ^, atf. atf. Am oa oaib ibfb fbme me,, a` eb ebs` s`eo eoba bamt mth h ia EIM as útb` útb` per pere lb lbma mas s ia ibekmöstbfh. Eiaoås, `e axtreffböm ia EIM as ume carreobamte bophrtemta am `e fbamfbe lhramsa pere `e bi biam amtb tblb lbfe fefb fböm öm ia bm bmib ibvb vbiu iuhs hs (phr (phr aj ajao aop` p`h, h, vbh`e vbh `eihr ihras, as, `eirhm `eirhmas, as, vïftbo vïftboes es ia eff effbia biamta mtas s h kuarres), iatarobmefböm ia petarmb mbi iei, biamtblbfefböm ia fuarphs, atf.

ia e`kumhsaste vbrus.bmlhroefböm @hs sakoamthskamâtbfe ia EIM qua  phrtem sa iamhobmem kamas.

Cey verbhs verbhs prhfai prhfaibo bobam bamths ths ibl iblara aramta mtas s per pere e `e praperefböm ia EIM kamöobfh. Yhihs fhobamzem fh fhm m e` e`ku kume me lhro lhroe e ia `b `bsb sbs s fa fa`u `u`e `er, r, sa saku kubi bie e ia iasp iasprh rhta taff ffbö böm m y rafu rafupa pare refb fböm öm ia EI EIM. M. @e @es s prbmfb prbmfbpe` pe`as as ibl iblara aramfb mfbes es amtra amtra ver verbhs bhs aml amlhqu hquas as reibfem am a` kreih ia iasprhtaffböm y am a` pash oh`afu`er oh`af u`er ia` EIM prhiu prhiufbih. fbih. @hs lefthras lefthras qua elaftem `hs oâthihs ia ebs`eobamth ia` EIM shm

Lbkures 5. Oh`âfu`e ia EIM.

`e aiei, `e luamta y a` teoeõh ia `e ouastre. @e prasamfbe ia prhtaïmes, `ïpbihs, ph`bsefårbihs, atf. atf. iu iure remt mta a `e prap praper eref efbö böm m ia ia`` EIM EIM pu puai aia a bmtarlarbr fhm `hs oâthihs ia emå`bsbs ia EIM e` raiuf raiufbr br `e fe fe`b `bie iei i ia` EIM. EIM. @h @hs s oâ oâth thihs ihs ia axtreffböm pere purblbfer albfezoamta a` EIM ia ibvarses luamtas ianam eieptersa am lumfböm ia lefthras fhoh a` teoeõh ia `e ouastre, `e lrasfure ia `e ouastre y a` fhmtambih nbhquïobfh ia `es fâ`u`es ia `es qua sa axtrea a` EIM. A` oâthih ia ebs`e ebs `eoba obamth mth ian iana a verber verber sakúm sakúm a` teoeõh teoeõh ia `e ouastre. @e lrasfure ia `e ouastre teonbâm bml`uya am `e tâfm tâfmbf bfe e ia ax axtr tref effb fböm öm.. @hs @hs oâ oâth thih ihs s ia axtref axt reffbö fböm m teo teonbâ nbâm m verïem verïem sakúm sakúm a` fhmtam fhmtambih bih nbhquïobfh ia `es fâ`u`es luamta. Whr ajaop`h, am a` fes esh h ia `e `es s neft neftar arbe bes, s, `hs prb prbmfbp mfbpe` e`as as nbhquïobf nbhqu ïobfhs hs prasa prasamtas mtas am um axtr axtrefth efth fa`u`er shm prhtaïmes, EIM y E^M. Whr `h temth, `e axtreffböm fhm lamh` h a` treteobamth fhm prhtaese, sakubih ia `e a`bobmefböm ia` E^M fhm rbnhmuf`aese, iaje ume oua ouastr stre e ia EI EIM M purh. purh. Asths Asths trete treteoba obamth mths s puaiam mh sar sulbfbamtas sb `es fâ`u`es teonbâm fh fhmt mtba bama mam m fe femt mtbi biei eias as sb sbkm kmbl blbf bfet etbv bves es ia htrh htrhs s nb nbh hquïobfhs. @h @hs s tajbihs vakate` e`a as shm pertbfu`eroamta iblïfb`as e asta raspafth ye qua e oamu oa muih ih fhmt fhmtba bama mam m krem kremia ias s femt femtbi biei eias as ia fernhc fer nhcbir bireth eths s qua mh sa a`b a`bob obmem mem oaibem oaibemta ta `e

Iamtrh ia `es lumfbhmas qua ``ave e fenh a` EIM phiaohs oamfbhmer: a` e`oefameobamth e `erkh p`ezh ia bmlhroefböm kamâtbfe, `e rap`bfefböm, as iafbr, `e fepefbiei ia cefar fhpbes ia sï obsoh, qu qua a `a pa paro robt bta a e `e bm bmlh lhro roef efbö böm m ka kamâ mâtb tbfe fe `e phsbnb phs bnb`bi `biei ei ia tremsl tremslarb arbrsa rsa ia ume fâ`u`e fâ`u`e e `es fâ fâ`u `u`e `es s cbje cbjes s y ia kama kamare refb fböm öm am ka kama mare refb fböm öm,, fhiblbfefböm ia `es prhtaïmes eiafueies pere feie fâ`u`e, fâ`u` e, bmtar bmtarvbama vbama am a` fhmtrh` fhmtrh` ia` oatenh`bsoh oatenh`bsoh fa`u`er oaibemta `e eyuie ia` E^M y oaibemta `e sïmtasbs ia prhtaïmes y chrohmes y phr ú`tboh as bophrtemta iastefer qua a` EIM bmtarvbama am `e outefböm. A` ebs`eobamth ia EIM sa ralbara e` prhfash ia axtr axtrear ear EI EIM M ia ume ume fâ fâ`u `u`e `e am lhro lhroe e pure pure.. @e axt xtrreffb effböm öm ia EIM as um pesh pesh pra` pra`bo bobbmer  mer  bophrtemta am a` qua sa hntbama EIM purblbfeih e pert pertbr br ia htrh htrhs s fh fhop ophm hmam amta tas s fa fa`u `u`e `era ras s fh fhoh oh prhtaïmes prht aïmes,, E^M y `ïpbihs. `ïpbihs. A` A` EIM sa puaia ebs`er  ia fue`qubar fâ`u`e muf`aeie ia ibvarses luamtas, temth vbves fhoh ouartes, fhoh semkra amtare, fena``h, fena` `h, aspar asparoe, oe, cuashs cuashs,, uões, tajbihs, cafas, p`uo p`uoes es ouiei ouieies es,, få fåsf sfer eres es ia cuav cuavh, h, se` se`bv bve, e, fâ`u`e fâ` u`es s apbta` apbta`be` be`as, as, hrbme, hrbme, nef neftar tarbes bes,, emboe` emboe`as. as. tajbihs h p`emtes. A` ebs ebs`eo `eobam bamth th ia EIM as maf mafase aserb rbh h per pere e ume verbaiei ia ep`bfefbhmas am fbamfbe, oaibfbme y

axtreffböm fhm lamh`.

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iuremta 57 obmuths y sa rafhkbö `e fepe efuhse suparbhr am um OFY muavh, iuremta a` prhfash mh sa oh` h`as asttö `e fepe epe bmtar mtaro oaib aibe, sa eõei eõeibbö f`hrhlhroh: e`fhch` bsheoï`bfh am ume prhphrfböm ia >9: >9: 5, sa oa oazf zf`ö `ö `a `amt mte e y fhop fhop``ateo ateoam amta ta bmvbrtbamih a` tunh, `uakh sa famtrblukh a` tunh e 5>777 rpo iuremta 57 obmuths, sa rafhkbö `e fepe efuhs efuhse e am um OF OFY Y muav muavh h y sa `a eõ eõei eibö bö 7,5

As bo boph phrt rtem emta ta oa oamf mfbh bhme mer, r, qu qua a `e `es s e` e`at ates es ia pasfeih shm ume luamta axfa`amta y fhmlben`a ia  EIM ia e`te fe`biei fhm ume sarba ia vamtejes. Am `e oeyhrïe ia `hs feshs, a` EIM sa hntbama ia `e rafh`affböm ia semkra ia `hs pafas. Am a` fesh ia aspafb asp afbas as rer reres es h am pa`bkr pa`bkrh h ia axtbmf axtbmfböm böm,, ast asta a oâthih mh as rafhoamien`a. Whr `h temth, a` tajbih ia `es e`ates sa use e oamuih pere `e axtreffböm ia EIM EIM phrqua phrqua a` ouastr ouastrah ah as ra` ra`etb etbveo veoamt amta a råpb rå pbih ih,, `hkï `hkïst stbf bfeo eoam amta ta sb sbop op`a `a y mh `a `ate te`. `. Qm paquaõ paq uaõh h trhzh trhzh ia tajbih tajbih prhphr prhphrfbh fbhme me sul sulbfb bfbamt amta a  EIM pere `e WF^ y `e ibkastböm ia rastrbffböm. @h oås bophrtemta as qua `e rafh`affböm ia ouastres ia e`ates ia pasfeih mh ieõe e `hs pafas. Aste as ume fhmsbiarefböm ouy bophrtemta, aspaf aspafbe be`o `oam amta ta ra`a ra`ave vemt mta a fh fhm m ra rasp spaf afth th e `e `es s aspafbas reres y hrmeoamte`as. @es ouastres ia e`ates ia pafas sa utb`bzem pere astuibhs neseihs am EIM shnra ibvarsbiei kamâtbfe, sbstaoes ia eperaeobamth y iatarobmefböm ia `e petarmbiei ia `e `es s ph phn` n`ef efbh bhma mas s ia pa pafa fas s fh fhm m ume ume e` e`ta tare refb fböm öm

vh`uoam ia efateth shibh y um vh`uoam bkue` ia atemh` atem h` ca`eihia(577 (577%), %), 6saO oazf` oazf`ö ö nbam `e sh`ufböm ceste hntamar a` saiboamth ia EIM. (Am asa ohoamth sa puih var `hs `hs kruphs ia EIM am a` tunh), oås eia`emta sa bmfunö a` OFY e ->7 · F iu iure remt mta a 5 ch chre re,, `u `uak akh h sa sefö sefö a` tunh tunh y sa famt famtrb rblu lukh kh e 5777 5777 rpo rpo iur iurem emta ta 57 obmu obmuth ths s y iafemter a` shnrameiemta (Atemh`). Iasp Ia spuâs uâs sa `e `evö vö a` tunh tunh fh fhm m atem atemh` h` e` ;7 ;7%, %, famtrblukemih e 5777 rpo iuremta 57 obmuths y `u `uak akh h sa prhf prhfai aibbö e ia iafe femt mter er a` at atem emh` h` fhm fhm fubi fubiei eih h sb sbm m pa pari riar ar a` sa saib iboa oamt mth. h. (Vb (Vb sa tuvh tuvh e`kume e`k umes s ib iblbf lbfu`t u`teia eias, s, sa utb utb`bz `bzö ö ume pbpate pbpate pere pere axtrear`h), @uakh @uakh sa safö e` ebra `hs kråmu`hs ia  EIM e taopareture eonbamta oemtambamih a` tunh

oïmboe.

enbarth y e su. vaz safö fhm ia lb`trh pepa` safemta. safemta Oåssaeia`emta sapepa` rasus rasuspamib pamibö ö a`/ saiboa sai boamth mth am 1717-577 577ΰ` ΰ` ia teopöm teopöm YA h ekue ekue iastb`eie trbp`a astârb` y sa `a ekrakö >,7 ΰ` ia ^Mese ^Mes e e 577 ΰ` ia sh`ufböm sh`ufböm ia EIM EIM,, sa oemt oemtuvh uvh `e ou ouas astr tre e e 6; · F iure iuremt mta a > ch chre res. s. (Wer (Were e purblbfefb purbl bfefböm). öm). A`af A`aftrhl trhlhrasb hrasbs s ia `e ouastre ouastre shnra ka` ia ekerhse e` 7,;%.y sa astboö `e fhmfamtrefböm ia EIM e >7 · F h `bhlb`bfa `e ouastre fhm mbtrökamh `ïqubih.

 

Lbkure >. Axtreffböm ia EIM ia `es e`ates ia pasfeih.

Oâthih axparboamte` Wrboarh sa axtrejh >7-577 ok ia e`ate ia pasfeih fhm um nbs nbstur turïï ast astârb ârb`` y ume umes s pbmzes pbmzes.. @uakh @uakh sa Chohkambzerhm `es e`ates am 1: >9: 5) y `uakh sa oazf`ö

^aeftvhs Mafaserbhs 

Yrbs F` 5 O (pC 3, 7) - >, 1 o`.

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AIYE 7,1 O (pC 3,7) - 17 o`.

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E^Mese pemfraåtfe - 1 ok.

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Wrhambesa D - (hn`bkehrbh).

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Nesa Yrbs - 57 oO.

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AIYE - 5 oO.

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Yeopöm ia axreffböm >77O@

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`amte y ob `amte obmuf mufbhs bhseoa eoamta mta oai oaibem bemta ta `e bmvars bmvarsböm böm rapa ra patb tbie ie ia` OFY, OFY, sa fa famt mtrb rblu lukh kh e 5>77 5>777 7 rpo rpo

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Núlar YA:

 

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Lamh`: F`hrhlhroh: E`fhch` bsheoï`bfh (17

^afhoamiefbhmas

o`) - (>1: >9: 5). 

F`hrhlhroh: e`fhch` bsheoï`bfh (17 o`) (93: >).

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;7%, 577% ia aemh` (ca`eih).

 

Efaeh ia shibh 6O.

  Ibsfusböm y rasu`teih 

Nbn`bhkrelïe OfDaa fDaa,, Y. & OfDaa fDaa,, J. (>75 >759). 9). @e @es s nesa nesas s oh`afu`eras ia `e vbie. OfK^E[-CB@@ BMYA^EOA^BFEME AIBYH^AV, V.E. ia F. \

Fhmf`usböm|

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