Clc genetics chapter 3

Chapter 3: basic principles of molecular marker DNA marker and some biotechnology techniques Part 7: DNA, PCR, Electrophoresis methods... Restriction fragment length polymorphisms RFLPs

Chapter 3: basic principles of molecular marker (DNA marker) and some

biotechnology techniques

Part 7: DNA, PCR, Electrophoresis methods

1 Restriction fragment length polymorphisms (RFLPs)

3 Random amplified polymorphic DNA (RAPDs)

2 Amplified fragment length polymorphisms (AFLPs)

4 Simple sequence repeats (Microsatellites - SSRs)

AGCT

Restriction

site

Single nu

Marker RFLP Restriction Fragment Length Polymorphisms

DNA separation by gel electrophoresis

- Small DNA fragments move further through the gel than large fragments

Marker

B

A B

A B Restriction fragment length polymorphism (RFLP)

A B

Simple sequence length polymorphism (SSLP)

A

B

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Q: How many bands would you see on a gel after digesting

human and honey bee mtDNA with the EcoR1 restriction enzyme? Hint: mtDNA is circular in both humans and honey bees.!

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• 

• 

Size sorted!

Hình: Marker RAPD

Marker Random Amplified Polymorphic Marker chỉ thị RADP

B

Digestion of DNA with two enzymes

Ligation of adapters to

fragment ends

Primers complementary to adapters and to 3’ region of some of the fragments

Marker

Marker chỉ thị AFLP (Amplified Fragment Length Polymorphisms)

• 

• 

• 

Microsatellite marker = SSR (simple sequence repeat) marker

Repeat unit - simple sequence (mono-, di-, tri-, tetranucleotides)

Hình: Đa hình đoạn lặp lại

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Q: What’s the DNA sequence?!

Dựa trên sự biến đổi (đột biến) về mức độ di truyền DNA

Part 7: DNA, PCR, Electrophoresis methods

Where is DNA in plant cell ?

DNA extraction

DNA extraction

- How to work with DNA ? (DNA is very small, it can not see and smell)

- But it is very easy to amplify from 1 copy to many

copies using PCR method (Poly Chain Reaction)

1st cycle 2nd cycle 3rd cycle 4th cycle

Target region between two primers

Hình: Cơ chế biến tính DNA

Primer Primer

Synthesis

PCR (Polymerase Chain Reaction)

Basis elements use for DNA amplification

DNA separation by gel electrophoresis

- Small DNA fragments move further through the gel than large fragments