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GETTING STARTED; SETTING UP SAMPLE DATABASE GOOGLE DOCS
- SET UP config.py:
Copy template (config.template.py) to a working config. At a bash prompt ($):
$ cp config.template.py config.py
Edit this config.py file to contain values as indicated for a google docs account you wish to use for sample tracking
Also supply a path for scratch file creation, and the path to your rtd scripts (this folder)
- SET UP google docs tables:
Run the included initialization script:
$ initialize_sample_DB.py
- POPULATE DB_library_data google doc
Individual IDs will be drawn from the combination of:
DB_library_data (associates an individual sample with its ligated adapter) and
DB_index_by_well (associates the adapter with the expected barcode sequence present in the read data)
Login to google docs with the account you provided in config.py (docs.google.com)
Open DB_library_data, and add records as follows:
So far the DB_library_data has the following field (column) headers:
sampleID pool adapter adaptersversion flowcell lane index
sampleid can be whatever you like, although should contain only alphanumerics, dash or underscore.
Pool is the size-selection grouping (samples in the same sizing lane have the same pool).
Adapter is the well (or tube #) of the adapter to which this sample's DNA is ligated,
adaptersversion indicates the set of adapters this well comes from.
Flowcell, lane and index come when the samples are sequenced, and are
matched to the options of the same names upon invocation of preprocess_radtag_lane.py
Note on relationship between DB_library_data and DB_index_by_well:
the column mapping between these sheets is as follows:
DB_library_data:"adapter" <-> DB_index_by_well:"well"
DB_library_data:"adaptersversion" <-> DB_index_by_well:"adaptersversion"
Thus, assigning a read barcode to an individual is the process of
retrieving all records from DB_library_data matching the
(flowcell,lane,[index]) specified, and mapping
DB_library_data:"sampleid" to DB_index_by_well:"idx" (the sequence to
match in read data) via the field mappings stated above.
Two example DB_library_data lines follow:
sampleid pool adapter adaptersversion flowcell lane index
CRL129 1 A1 rad48 100617 1
YS_E001 1 29 flex48 120330_SN343_0318_AC0LB5ACXX 5 11
In the first case, the adapter set was "rad48" which we have arrayed
in a microplate, so the adapter that got added in the ligation is
identified by its well coordinate in the plate (this just comes from
my lab notebook). This sample was sequenced in lane 1 of the flowcell
we've called 100617. Any other samples with pool=1, flowcell=100617,
lane=1 are assumed to have been size-selected in the same gel/pippin
lane.
In the second case, the adapter set is "flex48", and these are in
numbered tubes, so rather than "LetterNumber" well coordinates, this
just got a number (the tube number). Its pool is also 1, but pools
are only keyed within their flowcell/lane/[index] set, so this pool 1
is not the same as the previous pool 1.
Finally, note that the first sample had no index; this is fine, this
just means we didn't do illumina-style "third read" multiplexing for
that sample, whereas for the second example we did (using index #11)
Example data for DB_library_data can be found at:
https://docs.google.com/spreadsheet/ccc?key=0AnHEwF1NpAxDdFlXRXliTElXQzF6dkswZk5HenlTclE
- RUN preprocess_radtag_lane.py
we start with the s_X_[1|2]_sequence[_indexZ].txt[.gz]
fastq files and feed those to preprocess_radtag_lane.py (X=lane,
Z=index; both from DB_library_data), or alternatively with any file
named whatever you like, as long as flowcell, lane and optionally
index are supplied as options to preprocess_radtag_lane.
See preprocess_radtag_lane.py help:
$ preprocess_radtag_lane.py -h
- RUN rtd_run.py
using the .uniqued.gz files generated for each lane/index, invoke rtd_run.py per help:
$ rtd_run.py -h
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