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Amplification of Intermethylated Sites (AIMS)

Epigenetic differences arise during the lifetime of monozygotic twins. Mario F. Fraga, Esteban Ballestar, Maria F. Paz et al. Amplification of Intermethylated Sites (AIMS). Important terms. Isoschizomer - pair of restriction enzymes

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Amplification of Intermethylated Sites (AIMS)

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  1. Epigenetic differences arise during the lifetime of monozygotic twins.Mario F. Fraga, Esteban Ballestar, Maria F. Paz et al. Amplification of Intermethylated Sites (AIMS)

  2. Important terms • Isoschizomer- pair of restriction enzymes • One restriction enzyme recognizes unmethylated or methylated sites • The other recognizes the unmethylated sites • Hypermethylation-formation of a methylation band • Hypomethylation- is the loss of a methlaytion band

  3. History • In 2002, AIMS was developed by • Jordi Frigola, Maria Ribas, Rosa-Ana Risques and Miguel Peinado • How they developed AIMS • They took the CpG island amplification method and modified it so they could view global DNA methylation

  4. General view of methylation

  5. AIMS is used for • Looking at large amounts of DNA that have been hyper-methylated or hypo-methylated • These methylation patterns can be seen using AIMS

  6. When and why we use AIMS • To study methylation changes in • Humans (Cancer and aging) • Honeybees • Aphids • Methylation changes are thought to be the reason why some people get cancer and why some people develop diseases later in life.

  7. Drawbacks • You have to know what you are identifying when viewing a film • You need to know how genomic mapping works • Why it is useful • Makes a lot of products • Makes readable products

  8. One way to simplify • Due to a large number of products from AIMS, capillary electrophoresis (CE) is used to further simplify and pinpoint specific areas of methylation

  9. Diagram of AIMS Technique

  10. Initial Digestion: SmaI

  11. Second Digestion:

  12. Adaptors

  13. Restrained PCR and Products

  14. Results Roughly 600 AIMS bands were resolved in the gels.

  15. Results • Between 0.5% and 35% of the bands were different within MZ twin pairs

  16. AIMS Product Analysis

  17. AIMS Products Analysis

  18. AIMS Products Conclusions • Twin pairs with the most differential AIMS bands corresponded to MZ twins who were older, had spent less time of their lives together, or had different natural health-medical histories.

  19. Literature Cited • Photos: • Google.images • http://www.web-books.com/MoBio/Free/Ch7F2.htm • http://www.hgu.mrc.ac.uk/people/r.meehan_researchb.html

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