915
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Changes
Added new SHROB Genotyping protocol
==New Method==This method was a tetra-pair ARMS system designed using the tool at http://primer1.soton.ac.uk/primer1.html * SHROB Flanking 5': GGTTGTTTCTCAATGCAGATAGTAAATT* SHROB Inner Fwd: CCTGGACACTGTCACCTAATGATTAAA* SHROB Inner Rev : TCCATTCAATAACCAGATATAACAGAGTA* SHROB Flanking Rev: TAATACTTGTTAACATTCGAAGGGATTC There are two outer control primers and two inner primers, one of which (the Fwd primer) is specific for the mutation as shown below. There should always be a 248bp fragment, and one or both of the other fragments: {| class="wikitable"! style="font-weight: bold;" | Genotype! colspan="3" style="text-align: center; font-weight: bold;" | Bands|-| Wild-Type| 248| 184| |-| Mutant| 248| | 118|-| Heterozygote| 248| 184| 118|} ==Original Method==This is the older method, suggested by Takaya ''et al.'' ===Primers===
* SHROB-Fwd AGT GAA TGC TGT GCA GTC
* SHROB-Rev AAG GTT CTT CCA TTC AAT
===Reference===
From Takaya K, Ogawa Y, Hiraoka J, Hosoda K, Yamori Y, Nakao K, Koletsky RJ. Nonsense mutation of leptin receptor in the obese spontaneously hypertensive Koletsky rat. Nat Genet 14: 130–1, 1996. [10.1038/ng1096-130 doi]