Genotyping mIRE1b 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

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