Pollen dynamics: Difference between revisions
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** PCR with 375 x 20 SSR primers | ** PCR with 375 x 20 SSR primers | ||
** Score them and make an automated DB for analysis | ** Score them and make an automated DB for analysis | ||
** Make meaningful predictions for the Mulugu SSO | |||
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Revision as of 08:59, 25 March 2009
- Resources:
- Pollen flow in Eucalyptus grandis determined by paternity analysis using microsatellite markers
- Tree Genetics & Genomes (2008) 4:37–47
- Work plan: Pollen_Dynamics
- Logistics
- 15 Maternal/Paternal* 25 progenies = 375 plants to be analyzed
- DNA Isolation: 375 samples
- PCR reaction : 375 * 20 = 7500
- Gel Run: 7500/60 = 125 runs in acrylamide (30+30 samples)
- Requirements
- Trays
- Qiagen DNA isolation kit: 2 (each can handle 250 samples)
- Ampli Taq 2 vials
- High resolution Agarose: 100g
- Strategy
- Seeds sowing
- Leaf sample collection from parental trees (15) and seedlings after one and half month
- DNA isolation from 375 samples
- Take representative sample and screen the primers to select 20 good
- PCR with 375 x 20 SSR primers
- Score them and make an automated DB for analysis
- Make meaningful predictions for the Mulugu SSO