WHOLE GENOME AMPLIFICATION WGA

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WHOLE GENOME AMPLIFICATION (WGA)   MULTIPLE
DISPLACEMENT AMPLIFICATION (MDA)   Rolling
Circle   Molecular Staging
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WHY   Converts ng of DNA (5ng) into ug (gt25ug)
overnight   Now, extract lymphocytes store in
liquid Nitrogen immortalize at PD (75) grow
up cell lines re-extract DNA   Far
cheaper/easier and can work for samples not
immortalized.  
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• MDA AdvantagesMultiple Displacement
  Amplification (MDA) is a proprietary
  amplification technology that enables the first
  effective whole genome amplification method.MDA
  enables for the first timeWhole Genome
  Amplification with enhanced coverage and minimal
  bias.The growing trend to study disease and
  drug response at the molecular level has focused
  attention on DNA as a precious resource. MDA
  technology enables the first effective, whole
  genome amplification method (W.G.A.) with the
  potential to transform the current practice of
  sample preparation and sample archiving for
  genetic testing.The MSI MDA technology platform
  enables
• A virtually unlimited number of tests per sample
• Use of smaller samples, such as buccal swabs,
  mouthwash, and finger prick
• Use of DNA from alternative sources, such as
  biopsies and needle aspirates
• An increase in the practice of archiving DNA for
  additional testing or re-testing
• Amplification directly from whole blood and
  cells

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Practically

• Add DNA (blood,saliva, etc) into tube
• Add reaction master mix
• Incubate o/n at 30oC

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MDA is a substantial advancement over currently
available W.G.A. methods, which are based on PCR.
The MDA method rapidly amplifies the genome with
comprehensive loci coverage and minimal bias
between any loci, yielding 10kb fragments in a
simple, scalable reaction.Optimal coverage and
minimal biasComprehensive coverage of all
single-copy gene sequences with unprecedented low
bias offers researchers the confidence needed for
an expanded range of downstream
applications.AccuracyUse of enzyme with
proofreading capability yields amplified material
true to the original material. This enables
researchers to perform assays, such as SNP
assays, where accuracy is paramount.ScalableLar
ger quantities of DNA can be generated simply by
increasing the reaction volume. A 1ml reaction
gives the same DNA yield per µl as a standard
100µl reaction. This ability to carry out large
volume reactions enables large-scale studies such
as high-density genotyping.Sample prep step
eliminatedThe MDA whole genome amplification
method can be carried out directly on crude lysed
cells, whole blood, buccal swabs, and other
biological samples, eliminating the costly and
time-consuming DNA purification
step.Assay-readyContaminants are effectively
diluted out during the MDA process, replacing the
need for DNA purification steps and allowing
downstream applications, such as TaqMan, to be
carried out with no inhibitory effects.High
throughputIdeally suited for 96-well-plate
automated systems, with no centrifugation and/or
filtrations needed.Long FragmentsThe MDA
process generates DNA in excess of 10kb in
length, which enables traditional molecular
biology methods such as RFLP.TaqMan is a
registered trademark of Applied Biosystems.
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The polymerase is able to copy, prime, and recopy
any DNA target without the need to repeatedly
denature the template as required by PCR.The
exponential nature of the amplification process
is powerful enough to amplify DNA from even a
single cell up to a million-fold in only a few
hours.
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