Starch gel electrophoresis (Polemoniaceae)

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Method synopsis

Standard enzyme electrophoresis technique used to determine genotypes at the 6PGD-2 locus for parentage assignment. Distinguishes between homozygous medium (MM) and homozygous slow (SS) allelic variants.

Synthesized from method descriptions across 6 papers using this protocol.

Procedure from a recent peer-reviewed implementation

Steps below were extracted from the most recent peer-reviewed implementation of this protocol in the corpus — Foraging efficiency and size matching in a plant-pollinator communitiy: the importance of sugar content and tongue length (2019), Ecology letters. Implementations in other papers (listed below) may differ.

1. Store and transport seeds
   Seeds were stored without refrigeration and transported to California
   Quantities: At least 6 seeds analyzed per plantDuration: September 1981 transportConditions: Room temperature storage
   Equipment: storage containers
2. Prepare seeds for electrophoresis
   Untreated seeds were crushed in a drop of 0.1 M Tris-HCl pH 7.0 with 0.01 M dithiothreitol, crude extract absorbed in wicks of Whatman #3 filter paper
   Quantities: 0.1 M Tris-HCl, 0.01 M dithiothreitol, individual seed crushingDuration: Not specifiedConditions: pH 7.0
   Equipment: Whatman #3 filter paper, crushing equipment, Tris-HCl buffer, dithiothreitol
3. Perform electrophoresis
   Electrophoresis performed horizontally on 11.7% electrostarch gels cooled with ice. Used Tris-citrate/lithium-borate system for PGM, histidine-citrate system for MDH, and Tris-citrate gel buffer for other enzymes
   Quantities: 25 enzyme systems screened initially, 5 polymorphic loci identified, 11.7% electrostarch concentrationDuration: Not specified Cooled with ice, various buffer systems depending on enzyme

Typical Equipment

• starch-gel electrophoresis equipment

Output Measurements

• individual genotype
• allelic composition

Papers Using This Protocol (6)

Species Studied With This Protocol