To Do
- add +A onto end-repaired sample
- set up overnight ligation
+A addition
- end-repaired sample with Invitrogen PureLink PCR Purification Kit, eluting in 35 μl of buffer
- 17ng/ul, or about 4 pmol total DNA @ 200bp
DNA sample (32 ul)
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1 mM dATP (10 ul)
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10x Klenow buffer (5 ul)
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Klenow exo (3’ to 5’ exo minus) (3 ul)
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- Incubate for 30 minutes at 37°C.
- MinElute PCR Purification Kit to purify on one QIAquick MinElute column, eluting in 10 μl of EB.
Adapter ligation
DNA sample (10 ul)
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water (5 ul)
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2x ligation buffer (25 ul)
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adapters @ 15uM (5 ul)
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ligase (5 ul)
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