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Chunk #35 — METHODS — shRNA Design Rules

Source
lincRNAs act in the circuitry controlling pluripotency and differentiation.
Embedded
yes

Text

For each lincRNA we enumerated all 21-mer sub-sequences and scored them as follows: (i) A “clamp score” was computed by looking at the nucleotides at positions 18, 19, and 20. If all three positions contained an A/T it was assigned a score of 4, if two positions were A/T it was assigned a score of 1.5 and if one was A/T it was assigned a score of 0.8. We then looked at positions 16, 17, and 21 if all 3 were A/T it was assigned a score of 1.25, if 2 were A/T it was assigned a score of 1.1, and if 1 was A/T is was assigned a score of 0.8. The clamp score was computed as the product of these two scores. (ii) A “GC score” was computed by looking at the total GC percentage of the 21-mer sequence. If the sequence was <25% GC it was assigned a score of 0.01 if it was <55% it was assigned a score of 3, if it was <60% it was assigned a score of 1, and if >60% it