Genetics 3, Lecture 1

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The dideoxy sequence method can be automated meaning

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Illumina sequencing

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Explanation

https://www.youtube.com/watch?v=fCd6B5HRaZ8

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DNA placed on chip in many short segments
Sequence by synthesis
Forward and reverse strands are sequenced separately
Denatured, primers attach and one nucleotide binds to template at a time and is detached
Uses DNA polymerase and fluorescent tags

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Sequencers

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Match the text
Pyrosequencing
Minlon

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Miniature sequencer ($1000)/Portable/Whole genome in minutes, no dyes
Sequencing by synthesis-as a particular nucleotide is added, flash of light produced by pyrophosphate (Phosphodiester bond)/ Good for short sequences

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Diagnosis

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Explanation

Eg. Avian Influenza strains were identified with Sanger sequencing and next-gen sequencing in China •800 human cases of avian influenza A (H7N9) virus infection have been reported since early 2013. (WHO, 2016)•Sequence analysis shows a higher than normal mutation rate in human hosts in three areas that resulted in higher pathogenicity

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PCR steps

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Repeat the denaturation and annealing of primers
Repeat the denaturation and annealing of primers
Extend the primers with Taq DNA polymerase
Extend the primers with Taq DNA polymerase
Denature to single strands and anneal primers
Extend the primers with Taq DNA polymerase

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For PCR to be efficient, it requires...

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Denaturation happens at _ and double stranded DNA melts to ____

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94-96 degrees

Single Stranded DNA

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Annealing happens at and primers bind to

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50-65 degrees

complementary sequences

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Elongation or Extension happens at and DNA polymerase binds to

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72 degrees

the annealed primers and extends DNA at the 3' end of the chain

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qPCR definition

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Applications of qPCR

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Quantify
Measure

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amount of starting DNA of a particular sequence
expression of a gene by quantifying mRNA transcripts of a particular gene or allele

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What are the 2 essential biochemical features for synthesis of DNA by DNA polymerase?

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Both DNA sequencing & PCR require DNA synthesis

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Minimum requirements for DNA synthesis in vitro:

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Strategies for DNA Sequencing

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Genetics 3, Lecture 1

Question 1 of 1

Time Left

The dideoxy sequence method can be automated meaning

Select the correct answer(s).

(0/0)

Illumina sequencing

Match the text (click and drag)

(0/0)

Explanation

1

2

3

4

5

DNA placed on chip in many short segments

Sequence by synthesis

Forward and reverse strands are sequenced separately

Denatured, primers attach and one nucleotide binds to template at a time and is detached

Uses DNA polymerase and fluorescent tags

Sequencers

Match the text (click and drag)

(0/0)

Pyrosequencing

Minlon

Miniature sequencer ($1000)/Portable/Whole genome in minutes, no dyes

Sequencing by synthesis-as a particular nucleotide is added, flash of light produced by pyrophosphate (Phosphodiester bond)/ Good for short sequences

Diagnosis

Select the correct answer(s).

(0/0)

Explanation

PCR steps

Match the text (click and drag)

(0/0)

1

2

3

4

5

6

Repeat the denaturation and annealing of primers

Repeat the denaturation and annealing of primers

Extend the primers with Taq DNA polymerase

Extend the primers with Taq DNA polymerase

Denature to single strands and anneal primers

Extend the primers with Taq DNA polymerase

For PCR to be efficient, it requires...

Select the correct answer(s).

(0/0)

Denaturation happens at _____ and double stranded DNA melts to ________

Fill in the blanks

(0/0)

Annealing happens at______ and primers bind to ______

Fill in the blanks

(0/0)

Elongation or Extension happens at ______ and DNA polymerase binds to ______

Fill in the blanks

(0/0)

qPCR definition

Select the correct answer(s).

(0/0)

Applications of qPCR

Match the text (click and drag)

(0/0)

Quantify

Measure

amount of starting DNA of a particular sequence

expression of a gene by quantifying mRNA transcripts of a particular gene or allele

What are the 2 essential biochemical features for synthesis of DNA by DNA polymerase?

Select the correct answer(s).

(0/0)

Both DNA sequencing & PCR require DNA synthesis

Select the correct answer(s).

(0/0)

Minimum requirements for DNA synthesis in vitro:

Select the correct answer(s).

(0/0)

Strategies for DNA Sequencing

Select the correct answer(s).

(0/0)

Denaturation happens at _ and double stranded DNA melts to ____

Annealing happens at and primers bind to

Elongation or Extension happens at and DNA polymerase binds to