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

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Presentation on theme: "Nucleic Acids DHPLC Applications in Biomedicine Pooria Gill PhD of Nanobiotechnology In The Name of Allah."— Presentation transcript:

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

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

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 Sizing and Mutation Detection DNA DNASep Cartridge 4.6 mm x 50 mm, for variety of DNA applications including Sizing and Mutation Detection, load capacity of 2µg especially for Rapid Mutation Detection 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 for Oligo Purification work only NUC OligoSep® Cartridge 4.6 mm x 50 mm; for Oligo Purification work only for Oligo Purification NUC OligoSep Prep HC Cartridge 7.8 mm x 50 mm, for Oligo Purification, load capacity of 5000µg for variety of RNA applications 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

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10 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

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13 Thanks for Your Attentions Reference:


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