User:Anthony Salvagno/Notebook/Research/2011/02/24/New pALS Reverse Primer

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Reason

I'm constantly getting two PCR products and the reason is because there is a sequence that matches my previous reverse primer by 67%. So I've checked through my pALS sequence and came up with another sequence that has a decent melting temp and has less matches. Here the next best match is 60% which is good enough (the forward primer has a 60% match too and that yields one product so this should work).

Sequence

  • 5' - Digoxygenin - TTCGCTCCAAGCTGGGCTGTGTG

New Unzipping Adapter

I also put in for a new unzipping bottom adapter. The previous oligo was this:

Oligo #2: Bottom adapter 1a - Biotin
Sequence 5' to 3': GAGCGGATXACTATACTACATTAGAATTCAGAC
Modification: X = biotin-dT

So I am now replacing the X with a T and putting the biotin on the 5' end. If there is a nick, this should help with unzipping. I don't think there can not be a nick with the biotin here so we shall see.

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