State-of-the art*

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State-of-the art ???
Graham Ramsay, Nature Biotechnology 1998,1640
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?????????????????????
1.?????(??)??(Human genome project)?????,?(1997??)
???2???800,000?cDNA??(EST)?????,??2005?????? 2.?
???????????11????0.6-4.2Mb?????4.6Mb???13Mb???,??
71???6???0.2???????
??,??????????????????????????????,?????????????
????!
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???????????????? ??????????

• ???????
• ?????????????????
• ????????????
• ????????????

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????????????????

• ????(Gene chip) ??DNA??(DNA Chip)?????(Biological
  chip,???????????????),?????????????????,??????????
  ??,???????????????????????????????

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• ????????????????????????????????????????????
  ??????,??????????(light-directed in situ
  synthesis)????????????????????????????????,???????
  ?????????,????????????

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????????????????????

????????????Southern Blotting?Northern
Blotting??(??Affymetrix??Southern?????????),??????
???????,?????????????????????,?????????
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???????????
??
???
??
???
??????
1.??????????NC??Nylon? 2.???????????????????????
?,???????????
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??????????

• ??
• ????????????????(???????)?????????????????(??????
  ??)?
• ????????????????????????,???????????????????????

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• ??
• ?????????????(???????????)
• ?????????(??????????????)
• ????????????(??????????????????????)

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????????????

• ????????????????????????????,?????????????????????
  ??????Affymetrix?????,??????????????????????,?????
  ???????????,?????????,????????????????????,???????
  ?????????????????

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??????????????????
12
??
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???????????
????????(substrate)???????????????????????????
????????,?????????????????,???????????????????,???
???????????????????????????????(?????)?
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???????????????
??,???(Robert S. Matson)??????????????????????
???????????
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???????????????
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• ????
• ??????????(??????????)???????????????????????
• ??????(streptavidin)??????(????????lassamine
  ?phycoerythrin)????
• ?????????????(epifluorescence microscope)?????????
  ??????????????
• ??????????,???????????????????

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• ??????? Watson-Crick??????????(mismatch)????????
  ??????????,??,?????????????5-35???????,??????????
  ????,??????????????????????????????????

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(No Transcript)
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??????????

• ??????? (Light directed in situ synthesis)

????????????????(photolithography)?????????????
??????????????????????????????????????(electrical
circuits),?????????????????????????????
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• ????(???)
• ???????
• ????????(photo labile protecting group)?NH2?????
• ????????(mask)??????????,????????????????,????????
  ?????,?????NH2???
• ?????????????????????????????????????,????????????
  ???????,????????????????
• ???,????????????????????

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????????????
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???????(Off-chip synthesis)

• ??????????????????,????????DNA??????????????
  ???????????????(???Cartesian Technologies???PixSys
  NQ/PA????)???????????????????????????,???????2500
  spots/cm2(Yershov et al??????????20,000-30,000spot
  s/cm2)?

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?????????????????????
??????5,744,305?????????????400spots/cm2
???Affymetrix ????
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?????????????????
???????????????
??,?????????????????????????????????????
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???????????????

• ??,?????????????????
• ??????????
• ?????????

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?????????

• ??,Glenn McGall??????????????????????????,????
  ???,???????16,000 spots/cm2??(????106spots/cm2)?
• ????????,???????????????????????????,??????,???
  ???????????????,?????????????????????????,????????
  ???????,??????,???????????(signal-noise
  ratio),????????????????

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• ???????????????????,????????????????,???????????
  ?????????????????????,????????????????

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(No Transcript)
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• ?????????????????????????????,?????????????????
  ?????????
• ??5??,????????????????????????????,????????????
  ????????????????????????????????????,???????????,?
  ??????????????????????????????????????????????????
  ?????????

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• ???????????????

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