Lab 2: Digestion and Identification

Trait Associations for DS markers and Allele Sizes

SNPs detected with dCAPS primers are summarized in the table.

				Uncut Allele		Cut Allele
DS#	Enzyme	SNP	Trait Associations	Allele	Size	Allele	Size
1	BamHI	C/T	C – Low Weight, Short height	T	162	C	142
3	EcoRI	C/T	A – Low Snout ratio	T	152	C	129
5	PstI	G/A	A – High Weight, Tall height	A	134	G	114
7	SalI	C/T	T – Low Weight	T	186	G	166

General Digestion

Volume	Master Mix	Solution
7 μl	–	PCR product
1 μl	4 samples + 1	Enzyme
2 μl	4 samples + 1(2 μl)	dH2O

1. 1. Prepare master mixes for digestions
   2. Incubate samples at 37°C for 30 minutes (10 minutes if using Fast Digest enzyme)
   3. Run samples on 2.5% TBE gel
   4. Visualize Gel and enter data in table below

	Positive Control			Uncut			DogA		DogB
SNP	Exp	Obs		Exp	Obs		Bands present	Genotype	Bands present	Genotype
DS1	142			162
DS3	129			152
DS5	114			134
DS7	166			186

Review the photo library of dogs and use the dCAPS to forensically identify the dogs.

Adapted from

• Hultman, Keith and Mellgren, Eve (2014). Fetching SNPs: A Dog Genotyping Laboratory for Undergraduate Biology. Genetics Society of America Peer-Reviewed Education Portal (GSA PREP): 2014.001; doi: 10.1534/gsaprep.2014.001