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Old 01-12-2012, 10:57 AM   #1
jlove
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Default "With-Bead" SPRI Library Preps - anyone doing this?

Has anyone (besides Broad) tried doing this "with-bead" library prep method? It basically involves leaving the beads in the reaction tubes throughout the library prep protocol, and serially eluting but retaining the beads with the eluate for the next step.

http://www.ncbi.nlm.nih.gov/pubmed/21205303

It makes great sense - the beads don't interfere with any of the reactions.
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Old 04-03-2013, 07:34 AM   #2
garciad1
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We tried the on-bead prep kit from KAPA Scientific and it works beautifully.
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Old 04-05-2013, 06:34 AM   #3
chevrm
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It might be useful for extremely precious samples where very low inputs are required, but doesn't make much of a difference otherwise.
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Old 02-23-2015, 07:22 AM   #4
julielaliberte66
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Default "with bead" for cfDNA

It works really nicely. I used it with cell free DNA samples. I save some money by re-using the SPRI beads. I did not have to transfer my DNA in different tubes after each reaction, which maximized my output.
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Old 02-23-2015, 11:55 PM   #5
sfranzenburg
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I did it and "re-activated" the beads with self-made PEG buffer. Worked really well.
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Old 04-20-2016, 08:07 AM   #6
JBKri
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I am considering to use elements of the "with-bead" protocol for a PCR cleanup. In the elution step (bottom of circle in Fig2 in the Fisher et al paper), would it be safe to store the eluted DNA (with beads) for days-weeks in a fridge, or is there potentially a risk (like possiblity of rust??)? Could this mix safely be frozen? In Agencourt's protocol it says DO NOT FREEZE, but I don't know if that is because the beads would be harmed or if the PEG solution (which is not present in the eluate) would be affected, or something else.

Last edited by JBKri; 04-20-2016 at 08:16 AM.
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