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Old 04-15-2013, 08:24 AM   #1
seqgirl123
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Default Ampure XP beads cleanup and concentration

Does anyone know what ratio of Ampure XP beads:sample I must use to remove material from my library that is 150bp and below? My library is in 30ul of 1x TE.

Also has anyone ever concentrated their final library in a DNA Speedvac/concentrator unit and sequenced it with reason being the library did not produce enough yield but the speedvac step allowed it to. I'm curious if the sequencing of this concentrated library worked out well?
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Old 04-16-2013, 02:40 AM   #2
Zaag
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You could try google! 1.0-1.5 ratio should do it.

Speedvac does not increase yield, just concentration. You can use it but make sure your library is in H2O.
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Old 04-24-2013, 09:40 AM   #3
jwfoley
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Quote:
Does anyone know what ratio of Ampure XP beads:sample I must use to remove material from my library that is 150bp and below? My library is in 30ul of 1x TE.
Look like about 0.8X from this: http://core-genomics.blogspot.ca/201...eads-work.html. But do your own test to make sure.

Also note that what matters is the amount of buffer, not of beads. You can use a stingy amount of beads and bring up the rest with 20% PEG, 2.5M NaCl. KAPA includes this buffer with their library kit so you can put in beads once for the first enzymatic reaction and then keep reusing the same beads with the same samples.

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Also has anyone ever concentrated their final library in a DNA Speedvac/concentrator unit and sequenced it with reason being the library did not produce enough yield but the speedvac step allowed it to.
I haven't but Microcon columns are very easy and can get it down to 10-20 ÁL.
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