Pratica di Shadowing: High Resolution Melting (HRM) analysis made easy. - Impara a parlare inglese con i video

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High resolution melting
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or HRM is a very sensitive method to detect sequence variation
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in different samples up to a resolution of a single base pair without the need of sequencing.
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Here you see a wild type of DNA or a PCR product.
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After mutation you may get a homozygous mutant in
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which both the alleles are mutated or a heterozygous mutant in which a single allele is mutated.
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Now you want to check if mutation is there or not.
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HRM is based upon the principle of DNA dissociation with increase in temperature followed by detection by a fluorescent dye.
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The DNA denituration is based on the GC content.
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Higher the GC content, stronger will be the DNA bonds and higher will be the melting temperature to dissociate the two strands.
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In HRM, the detection is assisted by specific binding of a saturating fluorescent dye to the double-stranded DNA
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and measuring its fluorescence in real time on increasing the temperature gradually.
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For HRM, you need to PCR amplify your target gene locus to obtain a specific amplicon.
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Smaller the size of amplicon, better the results will be.
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Then you add a saturating fluorescent dye
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that binds to the dna only in the double stranded form
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then you proceed for hrm analysis in a real time machine here you gradually increase the temperature
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and fluorescence is measured simultaneously the plot shows fluorescence in the y-axis and temperature on the x-axis
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3 different curves are obtained for wild, homozygous and heterozygous mutant.
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Here you see as homozygous mutant had a GC mutation as seen in the previous slide its TM is higher.
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So the curve shifts towards the right.
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Heterozygous mutant on the other hand have a mutation in only one strand.
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So it has a base mismatch and hence the TM is reduced and the curve shifts towards the left.
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method is so sensitive that it detects the variation in the melting temperature up to 0.2 degree Celsius.
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HRM being a simple, economic, rapid and precise method of DNA variant detection, it can be used for mutant identification,
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SNP genotyping, DNA methylation by bisulfide sequencing, DNA fingerprinting and so on.
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This video has been made on a special request by Ms. Sahily Russell.
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Vocabolario e note di pronuncia per questa lezione

Questo video contiene 27 frasi e 373 parole da ripetere con lo shadowing. Il parlato dura 2:30. Chi parla mantiene un ritmo costante di circa 149 parole al minuto, comodo per lo shadowing. Solo il 66% delle parole rientra nelle 3.000 più comuni dell’inglese, quindi il lessico è impegnativo.

Vocaboli chiave di questo video

15 parole del video che vale la pena imparare, con pronuncia e significato:

ParolaPronunciaSignificato
mutant sostantivo/ˈmju(t)n̩(t)/mutante
strand sostantivo/ˈstɹænd/filo
mutation sostantivo/mjuˈteɪʃən/mutazione
sequence sostantivo/ˈsiː.kwəns/sequenza
curve sostantivo/kɚv/curva
detection sostantivo/dɪˈtɛk.ʃən/identificazione, investigazione
melt verbo/mɛlt/sciogliere, fondere
sensitive aggettivo/ˈsɛn.sɪt.ɪv/ricettivo, sensibile
obtain verbo/əbˈteɪn/ottenere
gradually avverbio/ˈɡɹæd͡ʒuəli/gradualmente
variation sostantivo/ˌvɛəɹiˈeɪʃn̩/variazione
detect verbo/dɪˈtɛkt/individuare, rilevare
axis sostantivo/ˈæksɪs/asse
assist verbo/əˈsɪst/assistere, aiutare
hence avverbio/ˈhɛns/da qui

Pronuncia a cui fare attenzione

  • I suoni “sh” e “zh”: mutation /mjuˈteɪʃən/, detection /dɪˈtɛk.ʃən/, variation /ˌvɛəɹiˈeɪʃn̩/, identification /aɪˌdɛn(t)ɪfɪˈkeɪʃn̩/, dissociate /dɪˈsoʊ.ʃi.eɪt/
  • Parole lunghe — attenzione all’accento: identification /aɪˌdɛn(t)ɪfɪˈkeɪʃn̩/, simultaneously /ˌsaɪmlˈteɪnɪəsli/, homozygous /ˌhəʊməʊˈzaɪɡəs/, dissociate /dɪˈsoʊ.ʃi.eɪt/, dissociation /dɪˌsoʊsiˈeɪʃən/

Come esercitarsi con questo video

  1. Ascolta tutto il video una volta senza parlare e annota le parole che non conosci.
  2. Fai shadowing frase per frase a velocità normale, ripetendo ognuna finché il tuo ritmo coincide con quello di chi parla.
  3. Registrati e confronta con l’originale, facendo attenzione a parole come mutant, strand, mutation.

Cos'è la tecnica dello Shadowing?

Shadowing è una tecnica di apprendimento delle lingue supportata da studi scientifici, originariamente sviluppata per la formazione dei traduttori professionisti e resa popolare dal poliglotta Dr. Alexander Arguelles. Il metodo è semplice ma potente: ascolti un audio in inglese di madrelingua e lo ripeti immediatamente ad alta voce — come un'ombra che segue il parlante con un ritardo di solo 1–2 secondi. A differenza dell'ascolto passivo o degli esercizi di grammatica, lo shadowing costringe il tuo cervello e i muscoli della bocca a elaborare e riprodurre simultaneamente i modelli di discorso reale. La ricerca dimostra che migliora significativamente la precisione della pronuncia, l'intonazione, il ritmo, il discorso connesso, la comprensione dell'ascolto e la fluidità del parlato — rendendolo uno dei metodi più efficaci per la preparazione alla prova di speaking dell'IELTS e per la comunicazione reale in inglese.

Tecnica dello shadowing: leggi la guida completa passo dopo passo →