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=== Sequenciación por hibridación ===
A ''[[secuenciación por hibridación]]'' é un método non encimático que usa unha [[micromatriz de ADN]]. AEtiquétase singlefluorecentemente poolun ofsó DNAconxunto whosede sequenceADNs iscuxa tosecuencia bese determinedpretende isdeterminar fluorescentlye labeledhibrídase andcunha hybridizedmatriz toque ancontén arraysecuencias containingcoñecidas. knownOs sequences.fortes Strongsinais hybridizationque signalsindican fromhibridación aprocedentes givendun spotdeterminado onpunto theda arraymatriz identifiesserven itspara sequenceidentificar ina thesúa DNAsecuencia beingno ADN que se está a sequencedsecuenciar.<ref>{{cite journal |author=Hanna GJ |title=Comparison of Sequencing by Hybridization and Cycle Sequencing for Genotyping of Human Immunodeficiency Virus Type 1 Reverse Transcriptase |journal=J. Clin. Microbiol. |volume=38 |issue=7 |pages=2715–21 |date=1 July 2000|pmid=10878069 |pmc=87006 |url=http://jcm.asm.org/cgi/pmidlookup?view=long&pmid=10878069 |author-separator=, |author2=Johnson VA |author3=Kuritzkes DR |display-authors=3 |last4=Richman |first4=DD |last5=Martinez-Picado |first5=J |last6=Sutton |first6=L |last7=Hazelwood |first7=JD |last8=d'Aquila |first8=RT }}</ref>
=== Sequenciación con espectrometría de masas ===
=== Sequencing with mass spectrometry ===
A [[Massespectrometría spectrometryde masas]] maypode beutilizarse usedpara todeterminar determineas DNAsecuencias sequencesde ADN. Matrix-assistedA laserespectrometría desorptionde ionizationmasas time-of-flightde massionización/desorción spectrometry,de orláser asistido por matriz ou [[Matrix-assisted laser desorption/ionization|MALDI-TOF MS]], hasfoi specificallyinvestigada beencomo investigatedmétodo asalternativo aná alternative[[electroforese]] methoden toxel gelpara electrophoresisvisualizar forfragmentos visualizingde DNA fragmentsADN. WithCon thiseste methodmétodo, DNAos fragmentsfragmentos generateddee byADN chain-terminationxerados sequencingpolas reactionsreaccións arede comparedsecuenciación byde massterminación ratherde thancadea bycompáranse size.pola Thesúa massmasa ofen eachvez nucleotidede ispolo differentseu fromtamaño. theA othersmasa andde thiscada differencenucleótido isé diferente e esta difeenza é detectable bypor massespectroscopía spectrometryde masas. Single-nucleotideAs mutationsmutacións indun asó fragmentnucleótido cannun befragmento morepoden easilyser detecteddetectadas withmáis MSdoadamente thancon byespectroscopía gelde electrophoresismasas aloneque só por electroforese en xel. A MALDI-TOF MS canpode moredetectar easilymáis detectfacilmente differencesas betweendiferenza RNAentre fragmentsfragmentos de ARN, sopolo researchersque mayos indirectlyinvestigdores sequencepoden DNAindirectamente withsecuenciar MS-basedo ADN por estes métodos baseados na espectroscopía de methodsmasas byconvertendo convertingprimeiro ito toADN RNAen firstARN.<ref>{{cite journal| title=Mass-spectrometry DNA sequencing| author= J.R. Edwards, H.Ruparel, and J. Ju| journal=Mutation Research| volume=573| issue=1–2| pages=3–12|year=2005| pmid=15829234| doi=10.1016/j.mrfmmm.2004.07.021}}</ref>
TheA highermaior resolutionresolución ofdos DNAfragmentos fragmentsde permittedADN byque MS-basedpermiten methodsos ismétodos ofbaseados specialna interestespectroscopía tode researchersmasas iné forensicde scienceinterese especial para os investigadores en ciencia forense, asxa theyque maypoden wishter toque finddetectar [[single-nucleotidepolimorfismo dun só nucleótido|polimorfismos dun só polymorphismsnucleótido]] inen humanmostras DNAde samplesADn tohumano identifypara individualsidentificar a individuos. TheseEstas samplesmostras maypoden beestar highlymoi degradeddegradadas, sopolo forensicque researchersos ofteninvestigadores preferforenses a miúdo prefiren examinar o [[mitochondrialADN DNAmitocondrial]] forpola itssúa highermaior stabilityestabilidade ande applicationsaplicacións forpara lineageestudos studiesde parentesco. MS-basedEn sequencingEEUU, methodsos havemétodos beende usedsecuenciación tobaseados compareen theespectroscopía sequencesde ofmasas humanforon mitochondrialutilizados DNApara fromcomparar samplesas insecuencias ado ADN mitocondrial humano de mostras da bse de datos do [[Federal Bureau of Investigation|FBI]] database <ref>{{cite journal|last=Hall|first=Thomas A.|coauthors=Budowle, Bruce; Jiang, Yun; Blyn, Lawrence; Eshoo, Mark; Sannes-Lowery, Kristin A.; Sampath, Rangarajan; Drader, Jared J.; Hannis, James C.; Harrell, Patina; Samant, Vivek; White, Neill; Ecker, David J.; Hofstadler, Steven A.|title=Base composition analysis of human mitochondrial DNA using electrospray ionization mass spectrometry: A novel tool for the identification and differentiation of humans|journal=Analytical Biochemistry|year=2005|volume=344|issue=1|pages=53–69|doi=10.1016/j.ab.2005.05.028}}</ref> ande fromde bonesósos foundatopados inen massfosas gravescomúns ofde Worldsoldados Warda I[[Primeira soldiersGuerra Mundial]].<ref>{{cite journal|last=Howard|first=R|coauthors=Encheva, V; Thomson, J; Bache, K; Chan, YT; Cowen, S; Debenham, P; Dixon, A; Krause, JU; Krishan, E; Moore, D; Moore, V; Ojo, M; Rodrigues, S; Stokes, P; Walker, J; Zimmermann, W; Barallon, R|title=Comparative analysis of human mitochondrial DNA from World War I bone samples by DNA sequencing and ESI-TOF mass spectrometry.|journal=Forensic science international. Genetics|date=2011 Jun 15|pmid=21683667|doi=10.1016/j.fsigen.2011.05.009|volume=7|issue=1|pages=1–9}}</ref>
EarlyOs chain-terminationmétodos andde terminación de cadea temperá e TOF MS methodsdemostraron demonstratedpoder readler lengthslonxitudes ofde upADn tode ata 100 basepares de pairsbases.<ref>{{cite journal|last=Monforte|first=Joseph A.|coauthors=Becker, Christopher H.|title=High-throughput DNA analysis by time-of-flight mass spectrometry|journal=Nature Medicine|date=1 March 1997|volume=3|issue=3|pages=360–362|doi=10.1038/nm0397-360}}</ref> ResearchersOs haveinvestigadores beennon unableforon toquen exceedde thissuperar averageeste readtamaño sizemedio de lectura; likeigual chain-terminationque sequencinga secuenciación de terminación de cadea por si alonesoa, MS-baseda DNAecuenciación sequencingde mayADN notbaseada bena suitableespectroscopía forde largemasas pode non ser axeitada para proxectos de secuenciadción ''de novo'' sequencing projectsgrandes. EvenAínda soasí, aun recentestudo studyrecente didutilizou useas thelecturas shortde sequencesecuencias readscurtas ande massa spectroscopyespectroscopía tode comparemasas single-nucleotidepara polymorphismscomparar inpolimorfismos pathogenicdun só nucleótido en cepas patóxenas de ''[[Streptococcus]]'' strains.<ref>{{cite journal|last=Beres|first=S. B.|coauthors=Carroll, R. K.; Shea, P. R.; Sitkiewicz, I.; Martinez-Gutierrez, J. C.; Low, D. E.; McGeer, A.; Willey, B. M.; Green, K.; Tyrrell, G. J.; Goldman, T. D.; Feldgarden, M.; Birren, B. W.; Fofanov, Y.; Boos, J.; Wheaton, W. D.; Honisch, C.; Musser, J. M.|title=Molecular complexity of successive bacterial epidemics deconvoluted by comparative pathogenomics|journal=Proceedings of the National Academy of Sciences|date=8 February 2010|volume=107|issue=9|pages=4371–4376|doi=10.1073/pnas.0911295107}}</ref>
=== MicrofluidicSecuenciación de Sanger sequencingmicrofluídica ===
{{main|Sanger sequencing}}
In microfluidic Sanger sequencing the entire thermocycling amplification of DNA fragments as well as their separation by electrophoresis is done on a single glass wafer (approximately 10 cm in diameter) thus reducing the reagent usage as well as cost.<ref>{{cite journal|last=Kan|first=Cheuk-Wai|coauthors=Fredlake, Christopher P.; Doherty, Erin A. S.; Barron, Annelise E.|title=DNA sequencing and genotyping in miniaturized electrophoresis systems|journal=ELECTROPHORESIS|date=1 November 2004|volume=25|issue=21–22|pages=3564–3588|doi=10.1002/elps.200406161}}</ref> In some instances researchers have shown that they can increase the throughput of conventional sequencing through the use of microchips.<ref>{{cite journal |author=Ying-Ja Chen, Eric E. Roller and Xiaohua Huang |title=DNA sequencing by denaturation: experimental proof of concept with an integrated fluidic device |journal=Lab on Chip |volume= 10|issue=10 |pages=1153–1159 |year=2010 |doi= 10.1039/b921417h |url=}}</ref> Research will still need to be done in order to make this use of technology effective.
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