cDNA synthesis and microarray hybridization

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cDNA synthesis and microarray hybridization

• Mod2 Day7
• Tuesday Oct 28th

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Notes from FNT

• About Figures Use every opportunity to be
  informative!
• Label strain names with relevant genotype
• Tell a little about what you did to get
  generate the data

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Notes from FNT
About calculation if you measured A260 0.429
using 1100 then A260 of undiluted 2 ug x
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Microarray cDNA synthesis, microarray hybrization
Double ug RNA?
Anneal capture sequence 1, 2 80 10 Extend
with RT, 42 1.5 hrs Destroy RNA NaOH, mix on
array
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Microarrays tomorrow
Wash off unhybridized cDNA Hybridize with Cy3,
Cy5 fluors Wash off uncaptured fluors Scan
Next time youll get a data file with info for
44K spots!
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While you wait

• Get to know what a data set will look like
• then
• Normalization--why and how we do it
• Background subtraction--what its for, what if
  you get a neg
• Log2 transformations--what its for and how we
  do it