Mapping Mutations in C. elegans - Caenorhabditis Elegans: Molecular Genetics and Development - page 15

2. Set up five reaction tubes for each lysate, one for each primer set ( Table I ):

Reagent Amount per reaction:

H 2 O 3.5 m L

50 mM MgCl 2 0.1 m L

Primer mix (6.25 pmol/ m L each) a 0.4 m L

Taq Master Mix (Promega 9PIM750) 5.0 m L

a

Except for primers o1295 and o1312, which should be at 3.12 pmol/ m L)

Table I
ARMS-PCR primers

Chromosome I = pkP1114
o1287 TTTCGGAATCAGTTTTATCTTGACGGAT

o1288 TCCAACGATGCTTCAACCGC

o1289 TGTTTCAATTCTACGAGGCAAGATATCG

o1290 TCATCCTTGTCCCAGACTACAATCTCAA

N2 product: 280 bp
CB4856 product: 200 bp
Chromosome II = pkP2149
o1273 AAAAAACTTGAAGGAGACCGAGACACAT

o1274 ACCATCGCATTTGCTTCAGATCAG

o1275 TTCTTCTCGTGAAACAGCTTCATCAACT

o1276 ATTTCATTTGCAGCTCGGTAACATTTTC

N2 product: 225 bp
CB4856 product: 350 bp
Chromosome III = ce3-171
o1311 GGTGTTGGACCAGCTTTTGCG

o1312 GGATTTGGCGGAATCTATCACCTG dilute 1:2 (125 uM);

o1313 CGACCTTCTGCGTCAGTATTGTTGA

o1314 AAAATTCGGCGGTCTTCATGAACTT

N2 product: 350 bp
CB4856 product: 450 bp
Chromosome IV = ce4-167
o1204 GCATTATTGTTTTCCGATTTTAAATGGTC

o1205 GCTCAGGACGATACATGTTTAAAATGGTA

o1206 TCACACGATACATCAAAGGAGTATCGTC

o1207 TTCAACAACATAAACGTTCTGTGGATTC

N2 product: 190 bp
CB4856 product: 175 bp
Chromosome V = ce5-15
o1295 TTTTTACTGTCAAATCGTCATGAGAGCT diluted 1:2 (125 uM)

o1296 AAGTACTGGCCAAATTTCAACGATCAAT

o1297 CCTCGTTGTTCGTTTCAGGAAATTATGT

o1298 GGGCATATTAATGATAAGGGTTGCAAAA

N2 product: 200 bp
CB4856 product: 350 bp