1. Nucleic Acids DHPLC Applications in Biomedicine
In The Name of Allah
Nucleic Acids DHPLC Applications in Biomedicine
Pooria Gill
PhD of Nanobiotechnology

2. Nucleic Acids DNA Linear RNA Nature-made Man-made Circular Vectors
Genomic
Cytoplasmic
rRNA
Man-made
mRNA
tRNA
Oligo-primer
ncRNA
Oligo-probe
tmRNA
Q-DNA
Circular
siRNA
Mitochondric
Organoplasmic

3. Applications
DHPLC Systems (e.g. WAVE Systems) can be used for a wide range of applications including:
Mutation Detection
Sizing
Oligonucleotide Purification and Analysis
Forensics
Microbial Analysis
RNA Isolation and Purification
Single Base Extension
Methylation

4. 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/Primer-based Amplification Technologies

5. 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

6. CARTRIDGE dependent Applications e.g. via WAVE Systems
DNA DNASep Cartridge 4.6 mm x 50 mm, for variety of DNA applications including Sizing and Mutation Detection, load capacity of 2µg
DNA 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 OligoSep® Cartridge 4.6 mm x 50 mm; for Oligo Purification work only
NUC OligoSep Prep HC Cartridge 7.8 mm x 50 mm, for Oligo Purification, load capacity of 5000µg
RNA 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

10. Chromatograms and pedigree of patients 3 and 16. A
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.

11. Example of Oligo Analysis by DHPLC

13. Thanks for Your Attentions Reference: