User:Cassandra M Barrett/Notebook/Open Chromatin/2015/10/26

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Blunt Site Oligos

Purpose: to design oligos for insertion into MV10 Xba site for LCR

Methods:

Design oligos to be annealed to one another and inserted into XbaI site on MV10 vector. Elements required:

  • sticky overhangs for XbaI cuts
  • SmaI (CCCGGG)blunt cut site sequence

Following oligos were designed, keeping in mind to avoid a frameshift

5' CT AGG CAG CCC GGG CTA CG 3'

5' CTA GCG TAG CCC GGG CTG C 3'

Oligos will be ordered from IDT and assembled similarly to the following: https://www.openwetware.org/wiki/User:Karmella_Haynes/Notebook/BioBrick_cloning/2013/03/20

Annealed oligos will be inserted into XbaI digested MV10 for subsequent LCR using SmaI for blunt digestion.

New ~40bp long LCR bridges were designed as well to be compatible with the new ends created by this insertion.