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FruitBreedomics – Technical SessionSampling procedures: Hands on experience and practical detailsI. Baumgartner (Agroscope)
San Giuseppe di Comacchio (Italy), June 19th 2015
Overview
• 96-well format – why?• Coding systems for plant identification• Demonstration on steps of leave sampling
procedure
Companies process samples in a 96-well format
Coding systems for individual plant identification
Individual labeling of seedlings• combination number• seedling number• (parents, resistances)
Positional coding system • 4 plates of 6 x 4 pots represent
96-well plate• No individual seedling labeling
1 2 3 4 5 6 7 8 9 10 11 12
A
B
C
D
E
F
G
H
1 - I 1 - II
1 - III 1 - IV
Positional coding system I
Plants in 4 plates of 6 x 4 pots Leave samples in 96-well plate
1 2 3 4 5 6 7 8 9 10 11 12
A
B
C
D
E
F
G
H
1 - I 1 - II
1 - III 1 - IV
1 2 3 4 5 6 7 8 9 10 11 12
A
B
C
D
E
F
G
H
2 - I 2 - II
2 - III 2 - IV
Positional coding system II
LGC leaf sampling I
LGC leaf sampling II
LGC leaf sampling III
• Plates of 6 x 4 pots allowed plant identification without labelling
• Coding system not easy for processing of plants
• Missing plants were replaced by living plants to fill up gaps in the plates (cost), laborious
• Puncturing the leaves and expedition of the plates was efficient
• The whole procedure was more time-consuming than expected
• Costs are relatively low, DNA extraction is the most expensive, data point to low price
• Close interaction with the company (LGC genomics) was required and successful
KASP - SNP Genotyping
MAB experiences
Thank you for your attention
powdery mildew
Agroscope good food, healthy environment