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Old 07-07-2009, 01:01 PM   #1
lmsiew
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Location: San Jose

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Default Buying reagents from outside source to replace the 454 DNA library kit

I heard some people just buy enzymes from outside source instead of buying the DNA library kit from 454. It is much cheaper that way. I would like to try that.
I was wondering if anyone have a list of where to get the enzymes and protocol for making the library since concentration of the reagents/enzymes might be different.
Any insight or helpful advice is highly appreciated. Thank you.
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Old 07-07-2009, 01:29 PM   #2
kmcarr
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NEB already has available a kit to prepare genomic DNA libraries for the Illumina Genome analyzer (http://www.neb.com/nebecomm/products/productE6000.asp) which replaces the Illumina kit at a fraction of the cost. You must supply your own adapters and primers; these can be ordered from Illumina (at great cost) or you can get them synthesized at IDT. We tested the NEB kit, using leftover adapters & primers from some Illumina kits, side by side with the Illumina kit on the same sample of genomic DNA and it worked just as well as the Illumina. The cost is $50/library prep (plus costs for adapters & primers) when purchasing the 10 prep kit or $40/library when purchasing the 50 prep kit. I doubt you could source the individual components much cheaper.

Reportedly NEB is planning on releasing kits for 454 library prep soon. Again, I imagine you will need to supply your own adapters and primers but the cost for these is marginal from some place like IDT.

Last edited by kmcarr; 07-07-2009 at 01:33 PM.
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Old 07-07-2009, 01:39 PM   #3
lmsiew
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Thanks for your reply. Yes, I will order own adapters and primers.
Any idea where to order the immobilization beads?
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