Reconstruction of germline sequences from alignment data¶
The CreateGermlines tool takes the individual segment alignment information for each sequence and reconstructs a full length germline sequence from the V(D)J reference sequences.
To reconstruct the germline, CreateGermlines trims V(D)J germline segments
and N/P regions by alignment length and concatenates them together. It puts Ns
in the untemplated N/P regions and optionally masks the D with Ns
-g dmask). CreateGermlines also looks for and
corrects cases where the alignment tool assigned the same part of the input sequence
to two different regions (eg, assigning the same nucleotides to N/P and J).
At the end of the germline reconstruction process, each sequence has been assigned a germline specific to the sequence.
When the (
--cloned) flag is specified, the
process is the same except it is clone specific and results in the
creation of one germline per clone. CreateGermlines selects first a
single V and J allele to use as the germline from all the assigned
annotations in each clone. The selection is made by simple majority rule of all
the allele calls in the clone. After the germline reconstruction process, all
sequences belonging to the same clone have been assigned the same germline.