Ron Lab


 

Genotyping mIRE1β K.O alleles


Primers

1. MIRE.i.1s:

GGA GAG ATT GGA GAA GCA CTA G (11/22)

2. PGK.UTR.5S:

CAG CTC ATT CCT CCA CTC ATG A (11/22)

3. MIRE.14AS:

AGC CCT GGA TAA GAG TCT GAA G (11/22)

Mutant allele

i.1S vs. UTR.5S size 231 nt

Wild-type allele

i.1S vs. 14AS size measured as ~450Bp

Worked out by Yuhong (August 31, 1998)

Buffer

10mM Tris pH=8.3, 50mM KCl, 1.5mM MgCl2

PCR Conditions

94˚(4 min) -> 35X [94˚(1 min), 55˚(10 min), 72˚(1 min)] -> 72˚(10 min)