DNA Methylation of Crassostrea gigas in Response to Ocean         AcidificationBy Andy Davison, Selina Cho, and Ying-     ...
Why do we care about Pacificoysters?
Why do we care about oceanacidification?
Why do we care aboutepigenetics?
Objectives   Quantify the physiological response of    diploid and triploid oysters exposed to    various acidified condi...
Methods
Restriction Enzyme Digestion
Methylated Cytosine Dot Blot
GelElectrophoresis
Results          Triploid and diploid oyster mortality for             control and high CO2 treatments
Restriction Enzyme DigestionsPCR                      All samples show elicit                      bands (~1140bp)        ...
Hsp70        Bands        appeared for        both MspI and        HpaII samples        Smaller, fainter        bands unde...
Quantitative PCR   control samples                   showed                   expression levels                   with a m...
Methylated Cytosine Dot Blot                                Methylation most common in both triploid                      ...
Conclusions   What went wrong?    ◦ Dot blot    ◦ Restriction enzymes   Diploids and Triploids are the same   Phenotypi...
Thank you
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FISH441: Oysters, acidification and methylation

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FISH441: Oysters, acidification and methylation

  1. 1. DNA Methylation of Crassostrea gigas in Response to Ocean AcidificationBy Andy Davison, Selina Cho, and Ying- Ying Lin
  2. 2. Why do we care about Pacificoysters?
  3. 3. Why do we care about oceanacidification?
  4. 4. Why do we care aboutepigenetics?
  5. 5. Objectives Quantify the physiological response of diploid and triploid oysters exposed to various acidified conditions Implications of varying levels of DNA methylation
  6. 6. Methods
  7. 7. Restriction Enzyme Digestion
  8. 8. Methylated Cytosine Dot Blot
  9. 9. GelElectrophoresis
  10. 10. Results Triploid and diploid oyster mortality for control and high CO2 treatments
  11. 11. Restriction Enzyme DigestionsPCR All samples show elicit bands (~1140bp) Row 2 sample 4- empty row Row 2 sample 17- same band patterns amp- gigasin 2
  12. 12. Hsp70 Bands appeared for both MspI and HpaII samples Smaller, fainter bands under only MspI samples
  13. 13. Quantitative PCR control samples showed expression levels with a median of 1.3e-07 hpaII digested samples- median of 2.5e-08 mspI digested samples- median 1.5e-08
  14. 14. Methylated Cytosine Dot Blot Methylation most common in both triploid and diploid oysters exposed to low pH conditions 14 of the 22 samples from the low pH treatment showed some level of methylation Four of the 22 samples from the dry treatment showed signs of methylation two of the 18 samples from the high pH, wet treatment demonstrated methylation Samples with 400ng of DNA showed methylation more frequently than 200ng samples All samples of triploid, low pH (400ng) showed methylation
  15. 15. Conclusions What went wrong? ◦ Dot blot ◦ Restriction enzymes Diploids and Triploids are the same Phenotypic Plasticity-stress response
  16. 16. Thank you

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