Difference between revisions of "SHROB Genotyping"

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(Made older method collapsable)
(added SOPs)
 
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==SOP==
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*[[Safety and Animal Training]]
  
 
==New Method==
 
==New Method==

Latest revision as of 17:09, 8 June 2020

SOP

New Method

This method was a tetra-pair ARMS system designed using the tool at http://primer1.soton.ac.uk/primer1.html

Primers

  • SHROB Flanking 5': GGTTGTTTCTCAATGCAGATAGTAAATT
  • SHROB Inner Fwd: CCTGGACACTGTCACCTAATGATTAAA
  • SHROB Inner Rev : TCCATTCAATAACCAGATATAACAGAGTA
  • SHROB Flanking Rev: TAATACTTGTTAACATTCGAAGGGATTC

Expected Results

There are two outer control primers and two inner primers, one of which (the Fwd primer) is specific for the mutation as shown below.

SHROB Genotyping.png

There should always be a 248bp fragment, and one or both of the other fragments:

Genotype Bands
Wild-Type 248 184
Mutant 248 118
Heterozygote 248 184 118
[Expand]

Original Method

This is the older method, suggested by Takaya et al.