In The Name of Allah
Nucleic Acids DHPLC
Applications in Biomedicine
Pooria Gill
PhD of Nanobiotechnology
pooriagill@yahoo.com
Nucleic Acids
 DNA
• Man-made
– Vectors
– Linear
• Nature-made
– Genomic
– Cytoplasmic
• Man-made
– Oligo-primer
– Oligo-probe
– Q-DNA
– Circular
• Nature-made
– Mitochondric
– Organoplasmic
 RNA
• Nature-made
– rRNA
– mRNA
– tRNA
– ncRNA
– tmRNA
• Man-made
– siRNA
Applications
 DHPLC Systems (e.g. WAVE Systems) can be
used for a wide range of applications including:
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Mutation Detection
Sizing
Oligonucleotide Purification and Analysis
Forensics
Microbial Analysis
RNA Isolation and Purification
Single Base Extension
Methylation
With the WAVE System you can:
 Scan fragments for both known and novel mutations/SNPs
without extensive resequencing
 Detect low-abundance mutations in heterogeneous samples
 Enrich for low-abundance alleles
 Fractionate complex mixtures of related fragments based on
differences in sequence content
 Perform targeted mutation/SNP scoring via primer extension
assays
 Analyze end-point products from allele-specific Probe/Primerbased Amplification Technologies
Probe/Primer-based Amplification Technologies
• Primer-based Amplifications:
– Non-Isothermal Amplifications e.g. PCR-based
Methods
– Isothermal Amplifications/Non-PCR-based
Amplifications:
– NASBA
– HDA
– SDA
– , and more than 45 N.A.I.A. Technologies
• Probe-based Amplifications:
– For example: Ligase Chain Reactions
CARTRIDGE dependent Applications e.g. via
WAVE Systems
 DNA-99-3510 DNASep Cartridge 4.6 mm x 50 mm, for variety of DNA
applications including Sizing and Mutation Detection, load capacity of 2µg
 DNA-99-3710 DNASep HT Cartridge 6.5 mm x 37 mm, for variety of DNA
application but especially for Rapid Mutation Detection, load capacity of 4µg
 NUC-99-3550 OligoSep® Cartridge 4.6 mm x 50 mm; for Oligo Purification
work only
 NUC-99-3860 OligoSep Prep HC Cartridge 7.8 mm x 50 mm, for Oligo
Purification, load capacity of 5000µg
 RNA-99-3810 RNASep® Prep Cartridge 7.8 mm x 50 mm, for variety of RNA
applications, load capacity of 50µg total RNA -OR- 1000ng of specific RNA
Chromatograms and pedigree of patients 3 and 16. A. DHPLC analysis in normal
control, patients 3 and 16. Both patients show abnormal DHPLC peak patterns with
two peaks in comparison to normal control. B. Sequencing data in patient 3 and his
family members. C. Sequencing data in patient 16 and his family members.
Example of Oligo Analysis by DHPLC
Thanks for Your Attentions
Reference: