16 April 2014

ddRADseq and restriction enzymes

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Wednesday April 16 notes

• Student Research Conference

• Fascinating discussion of Rmarkdown on ESS list

ddRADseq

AMPure Bead Purification of yesterday’s double-digest samples.

• added 75 ul AMPure Beads to the 50 ul rxn. Incubated 5 min room temp
• 2 min on magnetic plate. removed solution
• added 200 ul 70% ethanol (fresh). removed. repeat
• off plate, added 40 ul H₂O. 1 min
• on plate 2 min. transfered solution to new plate

Prep for ligation

• Remade P2 adapter as biotinylated DNA key for Streptavidin step
• 20 ul P2.1, 20 ul P2.2 (biotinylated), 10 ul 10x annealing buffer, 50 ul H₂O
• anneal 97.5°C for 2.5, cool to 21°C by stepping down 3°C every minute

ApTranscriptome

Meeting with SCH, NG, AN and MK to discuss thermal reactionome manuscript.