IGEM:MIT/2007/Notebook/2007-6-25

USING SEQUENCHER (Demo)

• Copy desired gene sequence (to check)

• Sequence tab
  • Create New Sequence
  • Give name
  • Paste sequence

• View tab
  • Translation – different reading frames (gives AAs)
  • Fifth button over – check presence of all common enzyme sites

• Cut map tab – show where different restriction sites
  • Select enzymes – look for different enzymes and click ok to see where that site is (Ensure presence/absence of restriction enzymes)
  • Bases

• View tab
  • Reverse and Com – gives reverse complementary sequence
  • Can look at sequencing data (ambiguities?)

• Take whole sequence (front, middle, back) and ensure all restriction enzymes are correct and present