Nucleic Acids DHPLCApplications in Biomedicine

Pooria GillPhD of Nanobiotechnology

pooriagill@yahoo.com

In The Name of Allah

Nucleic Acids DNA

– Linear

• Nature-made– Genomic

– Cytoplasmic

• Man-made– Oligo-primer

– Oligo-probe

– Q-DNA

– Circular

• Nature-made– Mitochondric

– Organoplasmic

• Man-made– Vectors

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:– 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/Primer-based 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 AttentionsReference: