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For target analysis of non conserved porcine miRNAs the tool Targetscan Custom was applied using the seed sequence of the identified miRNA.
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Anopheles miRNAs with a low level of sequence similarity were further used for the seed sequence alignment.
Furthermore we performed a functional analysis of the SOLiD shotgun sequences using the SEED classification [ 20], based on the given BLASTX output file.
Thus, using the seed and extend technique, only sequences sharing common kmers are considered for a possible alignment.
For seed finding we followed the same approach used to identify the seed sequence in microRNAs [ 15– 18], with short sequence windows sliding along the tRF sequence, without any location constraints.
Searches in the NCBI nr database using human LACTB as the seed sequence yielded more than five hundred bacterial proteins.
We performed homology search using iterative PSI-BLAST [ 29] using human DJ-1 as the seed sequence (NP_009193.2).
Using the identified seed sequences, we sought targets for the tRFs in the D. melanogaster genome based on perfect matches to 3'UTRs.
In the absence of comprehensive experimental miRNA targeting data, predictions for bound target mRNAs can be made using the miRNA seed sequences (Bartel 2009).
We construct an HMM for the seed sequence using the SAM modelfromalign software.
The seed sequence used for the initial HMM profile was that of the well-characterized PriA enzyme from S. coelicolor, referred to as PriA_Scoe [ 26, 28, 38, 42].
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