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By : Faizan Saleem
Introduction
 Multiple displacement amplification is a non-pcr based
amplification technique.
 It does not require multiple cycles for the amplification of
template DNA.
 An specific type of DNA polymerase is required i.e Φ29 DNA
polymerase.
 Even minute quantity of DNA from single cell can be amplified
with high fidelity rate.
 Whole amplification process requires a constant temperature of
30°C .
Φ29 DNA polymerase
 Φ29 DNA polymerase is obtained from bacteriophage Φ29.
 It has been extensively used in the field of molecular
biology.
 It comprises of two domains:
 A C-terminal domain (polymerase domain).
 An N-terminal domain (3'-5’ exonuclease domain).
 As compared to other type of polymerases, It has a higher
processitivity and proofreading ability.
How it Works:
Hyper Branched Structure
Pros and Cons
Pros :
 It provides higher processitivity , More than 70 kb of DNA
can be obtained.
 Higher fidelity rate, Φ29 DNA polymerase has a 3’–5'
proofreading activity that provides amplification error rate
to 1 in 106−107 bases compared to
conventional Taq polymerase.
 DNA from a single cell can be used for amplification.
 Does not require a thermo cycler .
Cons :
 Over amplification of a certain region may occur.
 Primer dimer formation is common occurrence.
 Allelic Dropout may occur in case of heterozygous sample.
 Requires constant temperature.
Applications of MDA
 Single cell genome sequencing.
 Forensic Analysis.
 SNP genotyping
 Southern Blotting etc.
Recent Work Done :
Multiple displacement amplification

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Multiple displacement amplification

  • 1. By : Faizan Saleem
  • 2. Introduction  Multiple displacement amplification is a non-pcr based amplification technique.  It does not require multiple cycles for the amplification of template DNA.  An specific type of DNA polymerase is required i.e Φ29 DNA polymerase.  Even minute quantity of DNA from single cell can be amplified with high fidelity rate.  Whole amplification process requires a constant temperature of 30°C .
  • 3. Φ29 DNA polymerase  Φ29 DNA polymerase is obtained from bacteriophage Φ29.  It has been extensively used in the field of molecular biology.  It comprises of two domains:  A C-terminal domain (polymerase domain).  An N-terminal domain (3'-5’ exonuclease domain).  As compared to other type of polymerases, It has a higher processitivity and proofreading ability.
  • 5.
  • 8. Pros :  It provides higher processitivity , More than 70 kb of DNA can be obtained.  Higher fidelity rate, Φ29 DNA polymerase has a 3’–5' proofreading activity that provides amplification error rate to 1 in 106−107 bases compared to conventional Taq polymerase.  DNA from a single cell can be used for amplification.  Does not require a thermo cycler .
  • 9. Cons :  Over amplification of a certain region may occur.  Primer dimer formation is common occurrence.  Allelic Dropout may occur in case of heterozygous sample.  Requires constant temperature.
  • 10. Applications of MDA  Single cell genome sequencing.  Forensic Analysis.  SNP genotyping  Southern Blotting etc.