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Equilibrate Beads to room Temperature
Add 24 ul of MagBeads to each well and 15 ul of replicate to same well of replicate
Pipette mix up and down 10 times.
Incubate at RT for 5 minutes
Secure plate on magnet plate; incubate at RT for 5 minutes (until wells are clear)
Remove 65 ul from each well; keep tips to left or right depending on the column to avoid bead pellet.
Add 100 ul Fresh 80% EtOH to each well. Incubate 30 seconds. Remove 100 ul from each well
Add 100 ul Fresh 80% EtOH to each well. Incubate 30 seconds. Remove 100 ul from each well
Reaspirate from each well to assure maximum EtOH removal
Allow plate to air dry for 7 minutes.
Remove sample plate from magnet plate.
Add 40 ul TE; pipette mix 10+ times. Incubate 2 minutes at RT.
Place sample plate back on magnet for 5 minutes or until all wells are cleared.
Transfer 40 ul to labeled transparent plate (Plate name_PCR_MIDs)
Store in -20 until ready to qPCR
TapeStation
Pool:
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Samples:
Pool, followed by Brock 2, column 2 (sample 5 is the blank)
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Blank:
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qPCR
45 nM
2.2 ul sample plus 97.8 ul RSB to make 1 nM
Sequence Dilution
Dilute to 1 nM:
2.2 ul sample plus 97.8 ul RSB
Add 10% PhiX and dilute to 65 pM:
13 ul sample plus 1.3 ul PhiX plus 185.7 RSB