User:Daniel A. Vargas/Notebook/General lab notebook/2015/08/04: Difference between revisions
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'''LCR Assembly''' | '''LCR Assembly''' | ||
Prep the Bridge Oligos | '''Prep the Bridge Oligos''' | ||
# MV10/Gal4DB bridge: LCRb_MV10tctGal4DB_rc | # MV10/Gal4DB bridge: LCRb_MV10tctGal4DB_rc | ||
# | # | ||
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PNK treatments | '''PNK treatments''' | ||
# MV10 + Gal4DB-mCh + ATF2 | # MV10 + Gal4DB-mCh + ATF2 | ||
# MV10 (neg ctrl.) | # MV10 (neg ctrl.) | ||
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LCR Reactions | '''LCR Reactions''' | ||
# MV10 + Gal4DB-mCh + ATF2 | # MV10 + Gal4DB-mCh + ATF2 | ||
# MV10 (neg. ctrl.) | # MV10 (neg. ctrl.) |
Revision as of 13:14, 4 August 2015
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ASSEMBLY of MV10, ATF2, and Gal4DB/mCherry
LCR Assembly Prep the Bridge Oligos
Note: Final conc. in LCR rxn. will end up being 30 nanomolar (nM) each
Overview: Mix double-stranded DNA fragments, treat with PNK to add phosphate groups
Set up the following in PCR tubes:
Master Mix (in 1.5 mL tube)
Add master mix into 10 μL PNK DNA
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