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ARRAYED PRIMER EXTENSION
(APEX) AND IT'S ADVANCES
APEX (Arrayed
Primer Extension) - a new resequencing method for rapid
identification of mutations that combines the efficiency of an microarray-based
assay (alternative to gel-based methods) with the Sanger sequencing.
Microarray - a microchip
(glass slide) containing thousand of DNA segments in an ordered array,
which allows the simultaneous analysis of thousands of genetic
markers.
About APEX:
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APEX is performed on a
two-dimensional array of oligonucleotides immobilized via 5’ end
on a glass surface (‘DNA chip’).
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DNA sample is
PCR amplified and annealed to the primers, which flank the
nucleotides to be identified.
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Four unique
fluorescently labeled terminator nucleotides are used by DNA
polymerase in primer extension reaction.
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As a result each
primer identifies one base in the target sequence.
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Covalent bonds between oligo and
dye terminator allow the slides to be stringently washed, thereby
improving the signal to noise ratio.
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The time required for the complete
APEX analysis including sample preparation is less than four hours.
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1. Up to
6000 known 25-mer oligos
are immobilized via 5’ end
on a
coated glass surface (‘DNA chip’). |
2.
Complementary fragment of
PCR amplified sample DNA is
annealed to oligos. |
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 |
3.
Template dependent single
nucleotide extension by DNA
polymerase. Terminator
nucleotides
are labeled with 4
different fluorescent dyes. |
4. DNA
fragments and unused dye
terminators are washed off.
Signal detection. |
Advantages
of APEX-based
genotyping:
-
A complete sequence analysis system
tailored for the customer's needs
-
parallel
analysis enables analysis of hundreds to thousands of SNP-s
/mutations in a single
reaction
-
DNA
polymerase and four dye
terminators ensure high allelic discrimination
-
Arrays can be designed to be either locus or
disease-specific
Asper Biotech detects the
signals by four-channel
microarray imaging and analysis system - Genorama®
QuattroImagerTM. International patent application is filed for
Genorama® QuattroImagerTM microarray imaging system.
For more information:
Frequently Asked Questions
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