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Old 01-05-2012, 01:58 AM   #1
gio5
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Default PCR primers+adaptors

Hi, I am working for my Ms thesis on a metagenetic study of the meiofauna and I am going to amplify the 18S region, then sequencing with 454. My question is if I should reshape primers with te A and B adaptors, and the MID tags before amplify my samples.
I read the B adaptor has a biotina than should not be a normal primer. In every works that I read they always amplify 18s with adaptors and mid tags referring as normal primers. Does anyone can help me?

All the best
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Old 01-05-2012, 03:56 AM   #2
TonyBrooks
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You shouldn't need modified primers for what you want to do. Just append the adapter sequences and MIDs to the amplification primers before ordering. The MID only goes on the A primer, so you'll need a different one for each sample plus a common reverse (same for every sample) with the B adapter sequence appended.
There's a tech note somewhere on the my454 website with the sequences and design strategy.
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Old 01-05-2012, 09:32 AM   #3
gio5
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Thanks a lot TonyBrooks!
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