Talk:20.109(S07): Start-up expression engineering

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# empty
# empty
# ladder (???)
# ladder (???)
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#  
+
# -template
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#
+
# +template
-
# red team -template, contaminated (?)
+
# red team -template, contaminated
# red team +template, good
# red team +template, good
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#
 
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Revision as of 13:31, 6 April 2007

On Day 1 of Module 3, you did a few things:

  1. Learned about the SAGA chromatin-remodeling complex and chose a non-essential sub-unit of the SAGA complex to delete
  2. Designed primers to amplify the URA3 sequence with flanking sequence from upstream and downstream of the sub-unit chosen for deletion (On Day 2: This DNA sequence will be able to get into the nucleus of the yeast cells we are transforming after we have made them "competent" for DNA uptake. Once in the nucleus, the flanking sequences with homology to upstream and downstream sequence of your SAGA sub-unit will allow your DNA product from PCR to integrate into the yeast genome, via homologous recombination, kicking out your SAGA sub-unit sequence and replacing it with URA3 sequence)

Image:Gel mod3day1 PCR products.JPG

Gel lane loading order

Top row, Tue/Thur Section Samples

  1. empty
  2. ladder (100bp)
  3. purple team 1
  4. purple team 2
  5. blue team -template, good
  6. blue team +template, good
  7. red team +template, good
  8. red team -template, good
  9. pink team -template, contaminated (?)
  10. pink team +template, good
  11. green team -template, good
  12. green team +template, good
  13. yellow team +template, good
  14. empty

Bottom row, Wed/Fri Section Samples

  1. empty
  2. ladder (???)
  3. -template
  4. +template
  5. red team -template, contaminated
  6. red team +template, good
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