Shadowing Practice: High Resolution Melting (HRM) analysis made easy. - Learn English Speaking with Video

Les maken...
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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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Woordenschat en spreektips bij deze les

Deze video bevat 27 zinnen en 373 woorden om na te spreken. Het gesproken deel duurt 2:30. De spreker praat in een gelijkmatig tempo van ongeveer 149 woorden per minuut, prettig om te shadowen. Slechts 66% van de woorden hoort bij de 3.000 meest gebruikte Engelse woorden, dus de woordenschat is pittig.

Belangrijke woorden in deze video

15 woorden uit de video die het leren waard zijn, met uitspraak en betekenis:

WoordUitspraakBetekenis
mutant zelfstandig naamwoord/ˈmju(t)n̩(t)/mutant
mutation zelfstandig naamwoord/mjuˈteɪʃən/mutatie
sequence zelfstandig naamwoord/ˈsiː.kwəns/volgorde, reeks
curve zelfstandig naamwoord/kɚv/curve, kromming
melt werkwoord/mɛlt/smelten, laten smelten
sensitive bijvoeglijk naamwoord/ˈsɛn.sɪt.ɪv/gevoelig, prikkelbaar
obtain werkwoord/əbˈteɪn/verkrijgen
gradually bijwoord/ˈɡɹæd͡ʒuəli/geleidelijk, allengs
variation zelfstandig naamwoord/ˌvɛəɹiˈeɪʃn̩/afwisseling, variatie
detect werkwoord/dɪˈtɛkt/detecteren
axis zelfstandig naamwoord/ˈæksɪs/as
assist werkwoord/əˈsɪst/assisteren, bijstaan
hence bijwoord/ˈhɛns/van hier, heen
rapid bijvoeglijk naamwoord/ˈɹæp.ɪd/snel, snelle
gene zelfstandig naamwoord/d͡ʒiːn/gen

Uitspraak om op te letten

  • De klanken “sh” en “zh”: mutation /mjuˈteɪʃən/, detection /dɪˈtɛk.ʃən/, variation /ˌvɛəɹiˈeɪʃn̩/, identification /aɪˌdɛn(t)ɪfɪˈkeɪʃn̩/, dissociate /dɪˈsoʊ.ʃi.eɪt/
  • Lange woorden — let op de klemtoon: identification /aɪˌdɛn(t)ɪfɪˈkeɪʃn̩/, simultaneously /ˌsaɪmlˈteɪnɪəsli/, homozygous /ˌhəʊməʊˈzaɪɡəs/, dissociate /dɪˈsoʊ.ʃi.eɪt/, dissociation /dɪˌsoʊsiˈeɪʃən/

Zo oefen je met deze video

  1. Luister de hele video één keer zonder te spreken en noteer de woorden die je niet kent.
  2. Spreek zin voor zin na op normale snelheid en herhaal elke zin tot je ritme gelijk is aan dat van de spreker.
  3. Neem jezelf op en vergelijk met het origineel; let daarbij op woorden als mutant, mutation, sequence.

Wat is de Shadowing-techniek?

Shadowing is een wetenschappelijk onderbouwde taalleermethode die oorspronkelijk is ontwikkeld voor professionele tolkentraining en gepopulariseerd door polyglot Dr. Alexander Arguelles. De methode is eenvoudig maar krachtig: je luistert naar native Engelse audio en herhaalt het onmiddellijk hardop — als een schaduw die de spreker volgt met slechts 1–2 seconden vertraging. In tegenstelling tot passief luisteren of grammaticadrills, dwingt shadowing je hersenen en mondspieren om echte spraakpatronen tegelijkertijd te verwerken en te reproduceren. Onderzoek toont aan dat het de uitspraaknauwkeurigheid, intonatie, ritme, verbonden spraak, luisterbegrip en spreekvaardigheid aanzienlijk verbetert — waardoor het een van de meest effectieve methoden is voor IELTS Speaking-voorbereiding en echte Engelse communicatie.

Shadowing-techniek: lees de volledige stap-voor-stap-gids →