IUBio

LYS2 mutants

Rich Buckholz rgb12955 at glaxo.com
Thu Mar 7 10:12:36 EST 1996


Solange,

Overlapping PCR should do it.

(x= any old sequence ‚LYS2, get creative, use a restriction site for 
later analysis!)

Round 1:

LYS 2 flank
      „35bp                                    LYS2 flank
     ------> 																		xxxxxxx------->
    -------|---------------LYS2---------------|---------
           <-------xxxxxxx                     <--------

Round 2:

------>
---------------xxxxxxx
															xxxxxxx---------------
																		           <-------

Trafo yeast with this PCR product:

    „35bp                „35bp
--------------xxxxxxx------------
--------------xxxxxxx------------

Good luck,
Rich






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