Hello,
I am interested in the comparison/respective advantages and weak points of the various adapters technologies, from the point of view of in-vitro genomic libraries construction and amplification, the avoidance of artifacts, and ease of preparation.
1°) anyone with a synthetic view on this (including the adapter technologies of Solexa, Illumina, Solid, etc...) ?
I am interested in the finicky details also (eventual exo protection, eventual 3'-end and/or 5'-end modification, etc...)
2°) about the Solid adaptors:(see also http://seqanswers.com/forums/showthread.php?t=588)
I have read in this forum (http://seqanswers.com/forums/showthread.php?t=198, sci_guy), that the solid adaptors are non-phosphorylated, so that P1-P1, P2-P2, or P1-P2 dimers are not formed at the ligation step; (by the way, are the 3'TT-overhangs efficient at avoiding ligation to genomic DNA ?)
Also, for the amplification steps, the targets of the primers are generated by nick translation.
Those features appears clever; Does this technique readily outperforms the competitor's adapters technologies ?
3°) Would someone have the detailled solid protocol for library prep at hand (under electronic form), that could be forwarded/downloaded/or pasted in this thread?
I am interested in the comparison/respective advantages and weak points of the various adapters technologies, from the point of view of in-vitro genomic libraries construction and amplification, the avoidance of artifacts, and ease of preparation.
1°) anyone with a synthetic view on this (including the adapter technologies of Solexa, Illumina, Solid, etc...) ?
I am interested in the finicky details also (eventual exo protection, eventual 3'-end and/or 5'-end modification, etc...)
2°) about the Solid adaptors:(see also http://seqanswers.com/forums/showthread.php?t=588)
I have read in this forum (http://seqanswers.com/forums/showthread.php?t=198, sci_guy), that the solid adaptors are non-phosphorylated, so that P1-P1, P2-P2, or P1-P2 dimers are not formed at the ligation step; (by the way, are the 3'TT-overhangs efficient at avoiding ligation to genomic DNA ?)
Also, for the amplification steps, the targets of the primers are generated by nick translation.
Those features appears clever; Does this technique readily outperforms the competitor's adapters technologies ?
3°) Would someone have the detailled solid protocol for library prep at hand (under electronic form), that could be forwarded/downloaded/or pasted in this thread?
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