Description Usage Arguments Value Author(s) References See Also Examples
Function readsamples() reads the output of read processing and mapping workflow which has to consist of 4 columns 1) RNAid, 2)Insert start, 3)Insert end, 4)Unique barcode count. It combines separate files coming from the same treatment (e.g. controls) and calculates estimated unique counts (EUCs) by either (a) keeping unique counts (euc="counts"), (b) using formula from Fu GK et al. PNAS 2011 (binomial distribution calculation) (euc="Fu") or (c) using method described in Kielpinski and Vinther, NAR 2014 (euc="HRF-Seq") If euc="Fu" then the count of all possible barcodes is required (m), e.g. if you use 7 nucleotide, fully degenerate random barcodes (NNNNNNN) then m=16384 (m=4**7) If euc="HRF-Seq" then the path to a precomputed k2n file is required (generate using k2n_calc() function)(default: "counts")
1 | readsamples(samples, euc = "counts", m = "", k2n_files = "")
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samples |
vector with paths to unique_barcodes files to be combined |
euc |
method of calculating estimated unique counts (default: "counts") |
m |
random barcode complexity (required if and only if euc="Fu") |
k2n_files |
vector with paths to k2n files corresponding to files given in samples (required if and only if euc="HRF-Seq"; order important!). Recycled if necessary |
euc_GR GRanges containing information: 1) seqnames (sequence name; RNAid) 2) Start, 3) End, 4) EUC value of a given fragment
Lukasz Jan Kielpinski, Nikos Sidiropoulos
Fu, G.K., Hu, J., Wang, P.H., and Fodor, S.P. (2011). Counting individual DNA molecules by the stochastic attachment of diverse labels. Proc Natl Acad Sci U S A 108, 9026-9031. Kielpinski, L.J., and Vinther, J. (2014). Massive parallel-sequencing-based hydroxyl radical probing of RNA accessibility. Nucleic Acids Res.
1 2 | write("DummyRNA\t1\t2\t3",file="dummy_unique_barcode")
readsamples(samples = "dummy_unique_barcode", euc = "counts")
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