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

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27 משפטים
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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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Vocabulary and speaking notes for this lesson

This video has 27 sentences and 373 words to shadow. The speech runs for 2:30. The speaker talks at a steady 149 words per minute, a comfortable pace for shadowing. Only 66% of the words are among the 3,000 most common in English, so the vocabulary is demanding.

Key vocabulary in this video

15 words from the video worth learning, with pronunciation and meaning:

WordPronunciationMeaning
mutant noun/ˈmju(t)n̩(t)/That which has mutated, with one or more new characteristics from a mutation.
strand noun/ˈstɹænd/Each of the strings which, twisted together, make up a yarn, rope or cord.
mutation noun/mjuˈteɪʃən/Any alteration or change.
sequence noun/ˈsiː.kwəns/A set of things next to each other in a set order; a series.
curve noun/kɚv/A bend, such as in a road.
detection noun/dɪˈtɛk.ʃən/The act or process of detecting, uncovering, or finding out, the discovery of something new, hidden, or disguised.
melt verb/mɛlt/To change (or to be changed) from a solid state to a liquid state, usually by a gradual heat.
sensitive adjective/ˈsɛn.sɪt.ɪv/Having the faculty of sensation; pertaining to the senses.
obtain verb/əbˈteɪn/To get hold of; to gain possession of, to procure; to acquire, in any way.
gradually adverb/ˈɡɹæd͡ʒuəli/In a gradual manner; making slow progress; slowly.
variation noun/ˌvɛəɹiˈeɪʃn̩/The act of varying; a partial change in the form, position, state, or qualities of a thing.
detect verb/dɪˈtɛkt/To discover or find by careful search, examination, or probing.
axis noun/ˈæksɪs/An imaginary line around which an object spins (an axis of rotation) or is symmetrically arranged (an axis of symmetry).
bind verb/ˈbaɪ̯nd/To tie; to confine by any ligature.
assist verb/əˈsɪst/To help.

Pronunciation to watch

  • The “sh” and “zh” sounds: mutation /mjuˈteɪʃən/, detection /dɪˈtɛk.ʃən/, variation /ˌvɛəɹiˈeɪʃn̩/, identification /aɪˌdɛn(t)ɪfɪˈkeɪʃn̩/, dissociate /dɪˈsoʊ.ʃi.eɪt/
  • Long words — get the stress right: identification /aɪˌdɛn(t)ɪfɪˈkeɪʃn̩/, simultaneously /ˌsaɪmlˈteɪnɪəsli/, homozygous /ˌhəʊməʊˈzaɪɡəs/, dissociate /dɪˈsoʊ.ʃi.eɪt/, dissociation /dɪˌsoʊsiˈeɪʃən/

How to practise with this video

  1. Listen to the whole video once without speaking and note the words you do not know.
  2. Shadow it sentence by sentence at normal speed, repeating each one until your rhythm matches the speaker.
  3. Record yourself and compare with the original, paying attention to words like mutant, strand, mutation.

מהי טכניקת השאדואינג?

שאדואינג היא טכניקת למידת שפה מבוססת מדע שפותחה במקור להכשרת מתורגמנים מקצועיים ופופולרית על ידי הפוליגלוט ד"ר אלכסנדר ארגואלס. השיטה פשוטה אך עוצמתית: אתה מקשיב לאודיו באנגלית ילידית וחוזר עליו בקול רם מיד — כמו צל העוקב אחרי הדובר עם עיכוב של 1–2 שניות בלבד. בניגוד להאזנה פסיבית או תרגילי דקדוק, שאדואינג מכריח את המוח והשרירים בפה לעבד ולשחזר דפוסי דיבור אמיתיים בו זמנית. מחקרים מראים שזה משפר משמעותית את דיוק ההגייה, האינטונציה, הקצב, הדיבור המחובר, הבנת השמיעה ושטף הדיבור — מה שהופך אותו לאחת השיטות היעילות ביותר להכנה לIELTS Speaking ולתקשורת באנגלית בעולם האמיתי.

טכניקת Shadowing: קרא את המדריך המלא שלב אחר שלב →