The researchers analyzed DNA from six different vegan meat and dairy substitutes using PCR and sequencing to check the accuracy of ingredient labels. All samples amplified with plant primers showed plant DNA, though two matched non-listed plant species. The mammalian primer produced plant DNA sequences from two samples, inconsistent with its intended use. Overall, the results supported accurate labeling for four products but found evidence against the labels of vegan hotdogs and cheese dip.

1. Abstract: Are vegan-labelled foods truly animal-free? Previous research has shown that some processed, packaged vegan foods have contained animal products. We investigated six different vegan meat and dairy
substitutes. We isolated DNA from the foods, and then used Polymerase Chain Reaction (PCR) to amplify a specific target gene. We used three primers on each sample: plant, fish, and mammal/insect. The resulting
gene copies were sequenced and then the sequence data was analyzed using bioinformatic techniques.
Acknowledgements: We would like to acknowledge Lansing Community College for funding the project, GENEWIZ, Inc. for providing DNA sequence
data, and the iPlant Collaborative and NCBI for providing bioinformatics tools. We would also like to acknowledge Cold Spring Harbor DNA Learning
Lab for experimental protocols.
DNA Analysis: Are Packaged Vegan Foods Really AnimalDNA Analysis: Are Packaged Vegan Foods Really Animal
Free?Free?
Shannon Best, Elise DesJardins, Ender Dettlaff, Sarah Hancock, Lurah Peterson, Eric Robins and Melinda Wilson.
Molecular Biotechnology Program, Lansing Community College, Lansing, MITGTTGGATTTAAAGCTGGTGTTAAGGATTATAAATTGACTTATTACACCCCGGAGTATGAAACCAAGGATACTGATATCTTGGCAGCATTCCGAGTAACTCCTCAGCCCGGGGTTCCGCCCGAAGAAGCAGGGGCTGCAGTAGCTGCCGAATCTTCTACTGGTACATGGACAACTGTTTGGACTGATGGACTTACCAGTCTTGATCGTTACAAAGGGCGATGCTATCACATCGAGCCCGTTGTTGGG
AGGAAAATCAATTTATCGCTTATGTAGCTTATCCATTAGACCTATTTGAAGAGGGTTCTGTTACTAACATGTTTACTTCCATTGTGGGTAACGTATTTGGTTTCAAAGCCCTACGCGCTCTACGTCTGGAGGATCTGCGAATTCCCCCTACTTATTCAAAAACTTTCCAAGGTCCGCCTCATGGTATCCAAGTTGAAAGGGATAAGTTGAACAAGTACGGCCGTCCTTTTTTGGGATGTACTATTAAACCA
AATTGGGAT
PlantDNABarcode
DNAIdentified
Sequence Variation
Consensus
Wheat
Cauliflower, Cabbage
Japanese Laurel
Soy
Pea
Pea
Plant Primer Mammal Primer Fish Primer
Conclusions
The purpose of this project was to determine if the
ingredients of vegan products are consistent with the
labeling. To do this we looked at DNA barcodes, gel
electrophoresis, and chromatogram data. Bands on all the
gels indicated that amplification of the DNA occurred with
all primers. The electrophoresis gel results for the plant
primer supported our expectations, that plant material is in
processed vegan foods. Although the fish and
mammal/insect primer gels indicated evidence of
mammalian/insect and fish DNA, no DNA sequence was
found to match mammalian/insect or fish DNA sequence.
Interestingly, the primers should have only amplified
mammal/insect and fish DNA during PCR, but the DNA
“barcoded” as plant species. The vegan bologna, Mozzarella,
hamburger, and chicken tenders all showed evidence of
having ingredients that are consistent with package labeling.
The vegan hotdogs and cheese dip showed evidence of
ingredients not on the label. In conclusion, our results
indicate that there was plant DNA but no mammal/insect or
fish DNA found in our vegan samples.
References
Cold Spring Harbor Laboratory. (2013). About DNA Subway. Retrieved (5 April 2016) from http://dnasubway.iplantcollaborative.org/about/
Lev. (2014, August 24). Common Food Product Certifications and Labeling Terms. Retrieved (5 April 2016) from https://www.recipal.com/blogs/72-
common-food-product-certifications-and-labeling-terms
Cold Spring Harbor Laboratory. (n.d) DNA Barcoding 101. Retrieved from http://www.dnabarcoding101.org/files/using-dna-barcodes.pdf
Rhoades, H. (2016, March 24). Growing Mustards: How To Plant Mustard Greens. Retrieved (14 April 2016) from
http://www.gardeningknowhow.com/edible/vegetables/greens/growing-mustard-greens.htm
CBOL (2010). Barcode of Life - Identifying Species with DNA Barcoding. Retrieved (5 April 2016) from
http://www.barcodeoflife.org/content/about/what-dna-barcoding
4
2
1
1
3
Results
DNA was analyzed from 18 PCR amplified vegan samples. All 6 samples
amplified with plant primers showed plant DNA. However, two of the
plant sample database search results matched plant species that were
not consistent with the product labels: hotdog DNA matched cabbage,
and chia cheese DNA matched a non-edible plant.
Using the mammalian primer, PCR amplified DNA sequence showed
matches to plant genes in vegan bologna and vegan mozzarella.
Although the results were consistent with the label, they were
inconsistent with using the mammalian primer.
Chromatogram of vegan mozzarella DNA sequence. This data shows distinct peaks which indicates DNA from a single species. Chromatogram of vegan bologna DNA sequence. This data shows multiple overlapping peaks which indicates DNA
from a mixture of species resulting in an unreliable barcode.
Plant
Primer
Fish
Primer
Mammal
& Insect
Primer
1
2
3
4
5
6
Agarose Gel Electrophoresis
Sample Name
Probe
Primer Species
E. Score Mismatch
Query
Length
% Identity
DNA
Purity QC
Plant
Triticum aestivum -
Wheat
0 0 1146 100%
Mammal Glycine max- Soy 0 21 556 96%
Fish Pisum saativum -Pea 1.00E-143 95 536 82%
Plant Glycine max - Soy 0 0 556 99%
Mammal
Brassica juncea -
Mustard greens
0 11 411 97%
Fish NR NR NR NR NR
Plant
Brassica oleracea -
Cabbage Family
0 4 1158 99%
Mammal NR NR NR NR NR
Fish
S.marmoratus -Rock
Fish 1.00E-18 7 71 Not Relevent
Plant
Aucuba japonica -
Japanese Laurel
0 29 572 95%
Mammal NR NR NR NR NR
Fish NR NR NR NR NR
Plant Lathyrus - Veiny pea 0 2 572 99%
Mammal NR NR NR NR NR
Fish NR NR NR NR NR
Plant Pisum saativum - Pea 0 2 1132 99%
Mammal NR NR NR NR NR
Fish NR NR NR NR NR
1.Vegan Bologna
2.Vegan
Mozzarella
3.Vegan Hotdogs
1.85
1.88
1.84
1.84
1.68
1.92
5.Vegan
Hamburger
4.Vegan Cheese
Dip
6.Vegan Chik'n
Tenders
Table 1: Vegan Sample Sequence Data Analysis Summary Using DNA Subway and GenBank Bioinformatic Tools.