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

11. Diagram of AIMS Technique

12. Initial Digestion: SmaI

13. Second Digestion:

14. Adaptors

15. Restrained PCR and Products

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

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

18. AIMS Product Analysis

19. AIMS Products Analysis

20. 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.

21. 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