3Holley’s method was distinct from Sanger’s. He began by isolating the alanine tRNA molecules and then cutting them into shorter pieces of unequal lengths, using enzymes. Then, he separated each fragment by size using column chromatography and ran various chemical techniques to figure out the sequence of each piece. Finally, he “aligned” these fragments by finding overlaps between them, thus reconstructing the full, 77-nucleotide strand.
def __init_subclass__[T](
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We even tried building hierarchies with 2-3 levels, but the number of shortcuts grew too fast for higher levels if we generated a full graph inside each cluster.