viral_seq 1.0.7 → 1.0.12

data/lib/viral_seq.rb

@@ -1,4 +1,4 @@
-# Copyright (c) 2019 Shuntai Zhou (shuntai.zhou@gmail.com)
+# Copyright (c) 2020 Shuntai Zhou (shuntai.zhou@gmail.com)
 #
 # Permission is hereby granted, free of charge, to any person obtaining a copy
 # of this software and associated documentation files (the "Software"), to deal
@@ -35,5 +35,8 @@ require_relative "viral_seq/seq_hash_pair"
 require_relative "viral_seq/sequence"
 require_relative "viral_seq/string"
 require_relative "viral_seq/version"
+require_relative "viral_seq/tcs_core"
+require_relative "viral_seq/tcs_json"
+
 
 require "muscle_bio"

data/lib/viral_seq/constant.rb

@@ -1,7 +1,11 @@
 module ViralSeq
-  
+
   # array for all amino acid one letter abbreviations
 
   AMINO_ACID_LIST = ["A", "C", "D", "E", "F", "G", "H", "I", "K", "L", "M", "N", "P", "Q", "R", "S", "T", "V", "W", "Y", "*"]
 
+  SDRM_HIV_PR_LIST = {}
+  SDRM_HIV_RT_LIST = {}
+  SDRM_HIV_IN_LIST = {}
+  
 end

data/lib/viral_seq/enumerable.rb

@@ -3,10 +3,6 @@
 #   array = [1,2,3,4,5,6,7,8,9,10]
 #   array.median
 #   => 5.5
-# @example sum
-#   array = [1,2,3,4,5,6,7,8,9,10]
-#   array.sum
-#   => 55
 # @example average number (mean)
 #   array = [1,2,3,4,5,6,7,8,9,10]
 #   array.mean
@@ -45,12 +41,6 @@ module Enumerable
     len % 2 == 1 ? sorted[len/2] : (sorted[len/2 - 1] + sorted[len/2]).to_f / 2
   end
 
-  # generate summed value
-  # @return [Numeric] summed value
-  def sum
-     self.inject(0){|accum, i| accum + i }
-  end
-
   # generate mean number
   # @return [Float] mean value
   def mean

data/lib/viral_seq/hash.rb

@@ -1,4 +1,4 @@
-# addition methods for Class::Hash required for ViralSeq
+# additional methods for Class::Hash required for ViralSeq
 
 class Hash
 

data/lib/viral_seq/hivdr.rb

@@ -1,6 +1,6 @@
 
 module ViralSeq
-  class SeqHash
+  class SDRM
 
     # functions to identify SDRMs from a ViralSeq::SeqHash object at HIV PR region.
     #   works for MPID-DR protocol (dx.doi.org/10.17504/protocols.io.useewbe)

data/lib/viral_seq/sdrm.rb

@@ -0,0 +1,43 @@
+module ViralSeq
+  class DRMs
+    def initialize (mutation_list = {})
+      @mutation_list = mutation_list
+    end
+
+    attr_accessor :mutation_list
+  end
+
+  def self.sdrm_hiv_pr(seq_hash)
+  end
+
+  def self.sdrm_hiv_rt(seq_hash)
+  end
+
+  def self.sdrm_hiv_in(seq_hash)
+  end
+
+  def self.list_from_json(file)
+  end
+
+  def self.list_from_csv(file)
+  end
+
+  def self.export_list_hiv_pr(file, format = :json)
+    if foramt == :json
+
+    end
+  end
+
+  def self.export_list_hiv_rt(file, format = :json)
+
+  end
+
+  def self.export_list_hiv_in(file, format = :json)
+
+  end
+
+  def drm_analysis(seq_hash)
+    mutation_list = self.mutation_list
+
+  end
+end

data/lib/viral_seq/seq_hash.rb

@@ -9,7 +9,7 @@ module ViralSeq
   #     # align with MUSCLE
   #   filtered_seqhash = aligned_pr_seqhash.hiv_seq_qc(2253, 2549, false, :HXB2)
   #     # filter nt sequences with the reference coordinates
-  #   filtered_seqhash = aligned_pr_seqhash.stop_codon[1]
+  #   filtered_seqhash = aligned_pr_seqhash.stop_codon[:without_stop_codon]
   #     # return a new ViralSeq::SeqHash object without stop codons
   #   filtered_seqhash = filtered_seqhash.a3g[1]
   #     # further filter out sequences with A3G hypermutations
@@ -130,8 +130,8 @@ module ViralSeq
           end
         end
       end
-      sequence_hash = Hash[*sequence_a]
-      quality_hash = Hash[*quality_a]
+      sequence_hash = Hash[sequence_a.each_slice(2).to_a]
+      quality_hash = Hash[quality_a.each_slice(2).to_a]
 
       seq_hash = ViralSeq::SeqHash.new
       seq_hash.dna_hash = sequence_hash
@@ -181,6 +181,7 @@ module ViralSeq
       new_seqhash = ViralSeq::SeqHash.new
       new_seqhash.dna_hash = self.dna_hash.merge(sh2.dna_hash)
       new_seqhash.aa_hash = self.aa_hash.merge(sh2.aa_hash)
+      new_seqhash.qc_hash = self.qc_hash.merge(sh2.qc_hash)
       new_seqhash.title = self.title + "_with_" + sh2.title
       new_seqhash.file = self.file + "," + sh2.file
       return new_seqhash
@@ -312,22 +313,22 @@ module ViralSeq
 
     # screen for sequences with stop codons.
     # @param (see #translate)
-    # @return [Array] of two elements [seqhash_stop_codon, seqhash_no_stop_codon],
+    # @return [Hash] of two SeqHash objects {with_stop_codon: seqHash, without_stop_codon: seqHash},
     #
-    #   # seqhash_stop_codon: ViralSeq::SeqHash object with stop codons
-    #   # seqhash_no_stop_codon: ViralSeq::SeqHash object without stop codons
+    #   # :with_stop_codon : ViralSeq::SeqHash object with stop codons
+    #   # :without_stop_codon: ViralSeq::SeqHash object without stop codons
     # @example given a hash of sequences, return a sub-hash with sequences only contains stop codons
     #   my_seqhash = ViralSeq::SeqHash.fa('my_fasta_file.fasta')
     #   my_seqhash.dna_hash
     #   => {">seq1"=>"ATAAGAACG", ">seq2"=>"ATATGAACG", ">seq3"=>"ATGAGAACG", ">seq4"=>"TATTAGACG", ">seq5"=>"CGCTGAACG"}
-    #   stop_codon_seqhash = my_seqhash.stop_codon[0]
+    #   stop_codon_seqhash = my_seqhash.stop_codon[:with_stop_codon]
     #   stop_codon_seqhash.dna_hash
     #   => {">seq2"=>"ATATGAACG", ">seq4"=>"TATTAGACG", ">seq5"=>"CGCTGAACG"}
     #   stop_codon_seqhash.aa_hash
     #   => {">seq2"=>"I*T", ">seq4"=>"Y*T", ">seq5"=>"R*T"}
     #   stop_codon_seqhash.title
     #   => "my_fasta_file_stop"
-    #   filtered_seqhash = my_seqhash.stop_codon[1]
+    #   filtered_seqhash = my_seqhash.stop_codon[:without_stop_codon]
     #   filtered_seqhash.aa_hash
     #   {">seq1"=>"IRT", ">seq3"=>"MRT"}
 
@@ -342,12 +343,15 @@ module ViralSeq
       seqhash1.title = self.title + "_stop"
       keys2 = aa_seqs.keys - keys
       seqhash2 = self.sub(keys2)
-      return [seqhash1, seqhash2]
+      return {
+        with_stop_codon: seqhash1,
+        without_stop_codon: seqhash2
+      }
     end #end of #stop_codon
 
 
     # create one consensus sequence from @dna_hash with an optional majority cut-off for mixed bases.
-    # @param cutoff [Float] majority cut-off for calling consensus bases. defult at simple majority (0.5), position with 15% "A" and 85% "G" will be called as "G" with 20% cut-off and as "R" with 10% cut-off.
+    # @param cutoff [Float] majority cut-off for calling consensus bases. defult at (0.5), position with 15% "A" and 85% "G" will be called as "G" with 20% cut-off and as "R" with 10% cut-off. Using (0) will return use simply majority rule (no cutoff)
     # @return [String] consensus sequence
     # @example consensus sequence from an array of sequences.
     #   seq_array = %w{ ATTTTTTTTT
@@ -379,11 +383,18 @@ module ViralSeq
         base_count = all_base.count_freq
         max_base_list = []
 
-        base_count.each do |k,v|
-          if v/seq_size.to_f >= cutoff
-            max_base_list << k
+        if cutoff.zero?
+          max_count = base_count.values.max
+          max_base_hash = base_count.select {|_k,v| v == max_count}
+          max_base_list = max_base_hash.keys
+        else
+          base_count.each do |k,v|
+            if v/seq_size.to_f >= cutoff
+              max_base_list << k
+            end
           end
         end
+
         consensus_seq += call_consensus_base(max_base_list)
       end
       return consensus_seq
@@ -394,14 +405,14 @@ module ViralSeq
     #   # control pattern: G[YN|RC] -> A[YN|RC]
     #   # use the sample consensus to determine potential a3g sites
     #   # Two criteria to identify hypermutation
-    #   # 1. Fisher's exact test on the frequencies of G to A mutation at A3G positons vs. non-A3G positions
+    #   # 1. Fisher's exact test on the frequencies of G to A mutation at A3G positions vs. non-A3G positions
     #   # 2. Poisson distribution of G to A mutations at A3G positions, outliers sequences
     #   # note:  criteria 2 only applies on a sequence file containing more than 20 sequences,
     #   #        b/c Poisson model does not do well on small sample size.
-    # @return [Array] three values.
-    #   first value, `array[0]`: a ViralSeq:SeqHash object for sequences with hypermutations
-    #   second value, `array[1]`: a ViralSeq:SeqHash object for sequences without hypermutations
-    #   third value, `array[2]`: a two-demensional array `[[a,b], [c,d]]` for statistic_info, including the following information,
+    # @return [Hash] three paris.
+    #   :a3g_seq: a ViralSeq:SeqHash object for sequences with hypermutations
+    #   :filtered_seq : a ViralSeq:SeqHash object for sequences without hypermutations
+    #   :stats : a two-demensional array `[[a,b], [c,d]]` for statistic_info, including the following information,
     #     # sequence tag
     #     # G to A mutation numbers at potential a3g positions
     #     # total potential a3g G positions
@@ -412,17 +423,17 @@ module ViralSeq
     # @example identify apobec3gf mutations from a sequence fasta file
     #   my_seqhash = ViralSeq::SeqHash.fa('spec/sample_files/sample_a3g_sequence1.fasta')
     #   hypermut = my_seqhash.a3g
-    #   hypermut[0].dna_hash.keys
+    #   hypermut[:a3g_seq].dna_hash.keys
     #   => [">Seq7", ">Seq14"]
-    #   hypermut[1].dna_hash.keys
+    #   hypermut[:filtered_seq].dna_hash.keys
     #   => [">Seq1", ">Seq2", ">Seq5"]
-    #   hypermut[2]
+    #   hypermut[:stats]
     #   => [[">Seq7", 23, 68, 1, 54, 18.26, 4.308329383112348e-06], [">Seq14", 45, 68, 9, 54, 3.97, 5.2143571971582974e-08]]
     #
     # @example identify apobec3gf mutations from another sequence fasta file
     #   my_seqhash = ViralSeq::SeqHash.fa('spec/sample_files/sample_a3g_sequence2.fasta')
     #   hypermut = my_seqhash.a3g
-    #   hypermut[2]
+    #   hypermut[:stats]
     #   => [[">CTAACACTCA_134_a3g-sample2", 4, 35, 0, 51, Infinity, 0.02465676660128911], [">ATAGTGCCCA_60_a3g-sample2", 4, 35, 1, 51, 5.83, 0.1534487353839561]]
     #   # notice sequence ">ATAGTGCCCA_60_a3g-sample2" has a p value at 0.15, greater than 0.05,
     #   # but it is still called as hypermutation sequence b/c it's Poisson outlier sequence.
@@ -515,7 +526,10 @@ module ViralSeq
       hm_seq_hash.title = self.title + "_hypermut"
       hm_seq_hash.file = self.file
       filtered_seq_hash = self.sub(self.dna_hash.keys - hm_hash.keys)
-      return [hm_seq_hash, filtered_seq_hash, hm_hash.values]
+      return { a3g_seq: hm_seq_hash,
+               filtered_seq: filtered_seq_hash,
+               stats: hm_hash.values
+              }
     end #end of #a3g_hypermut
 
     alias_method :a3g, :a3g_hypermut
@@ -535,7 +549,7 @@ module ViralSeq
       if sequences.size == 0
         return 0
       else
-        cut_off = 1
+        cut_off = Float::INFINITY
         l = sequences[0].size
         rate = sequences.size * error_rate
         count_mut = variant_for_poisson(sequences)
@@ -544,7 +558,7 @@ module ViralSeq
 
         poisson_hash.each do |k,v|
           cal = l * v
-          obs = count_mut[k] ? count_mut[k] : 0
+          obs = count_mut[k] ? count_mut[k] : 1
           if obs >= fold_cutoff * cal
             cut_off = k
             break
@@ -729,6 +743,7 @@ module ViralSeq
 
       seq_hash_unique.each do |seq|
         loc = ViralSeq::Sequence.new('', seq).locator(ref_option, path_to_muscle)
+        next unless loc # if locator tool fails, skip this seq.
         if start_nt.include?(loc[0]) && end_nt.include?(loc[1])
           if indel
             seq_hash_unique_pass << seq
@@ -1144,6 +1159,27 @@ module ViralSeq
       return new_sh
     end
 
+    # trim dna sequences based on the provided reference coordinates.
+    # @param start_nt [Integer,Range,Array] start nt position(s) on the refernce genome, can be single number (Integer) or a range of Integers (Range), or an Array
+    # @param end_nt [Integer,Range,Array] end nt position(s) on the refernce genome,can be single number (Integer) or a range of Integers (Range), or an Array
+    # @param ref_option [Symbol], name of reference genomes, options are `:HXB2`, `:NL43`, `:MAC239`
+    # @param path_to_muscle [String], path to the muscle executable, if not provided, use MuscleBio to run Muscle
+    # @return [ViralSeq::SeqHash] a new ViralSeq::SeqHash object with trimmed sequences
+
+    def trim(start_nt, end_nt, ref_option = :HXB2, path_to_muscle = false)
+      seq_hash = self.dna_hash.dup
+      seq_hash_unique = seq_hash.uniq_hash
+      trimmed_seq_hash = {}
+      seq_hash_unique.each do |seq, names|
+        trimmed_seq = ViralSeq::Sequence.new('', seq).sequence_clip(start_nt, end_nt, ref_option, path_to_muscle).dna
+        names.each do |name|
+          trimmed_seq_hash[name] = trimmed_seq
+        end
+      end
+      return_seq_hash = self.dup
+      return_seq_hash.dna_hash = trimmed_seq_hash
+      return return_seq_hash
+    end
 
     # start of private functions
     private

data/lib/viral_seq/seq_hash_pair.rb

@@ -80,6 +80,12 @@ module ViralSeq
       alias_method :fa, :new_from_fasta
     end
 
+    # the size of nt sequence hash of the SeqHashPair object
+    # @return [Integer] size of nt sequence hash of the SeqHash object
+    def size
+      self.dna_hash.size
+    end
+
     # Pair-end join function for KNOWN overlap size.
     # @param overlap [Integer] how many bases are overlapped. `0` means no overlap, R1 and R2 will be simply put together.
     # @param diff [Integer, Float] the maximum mismatch rate allowed for the overlapping region. default at 0.0, i.e. no mis-match allowed.
@@ -211,7 +217,7 @@ module ViralSeq
     # {minimal overlap set to 4. }
     def overlap_matrix(sequence1, sequence2)
       min_overlap = 4
-      max_overlap = [sequence1.size, sequence2.size].max
+      max_overlap = [sequence1.size, sequence2.size].min
       matrix_hash = {}
       (min_overlap..max_overlap).each do |overlap|
         matrix_hash[overlap] = sequence1[-overlap..-1].compare_with(sequence2[0, overlap])

data/lib/viral_seq/tcs_core.rb

@@ -0,0 +1,305 @@
+module ViralSeq
+
+  # Core functions for `tcs` pipeline
+
+  class TcsCore
+    class << self
+
+      # methods to calculate TCS consensus cut-off based on the maximum numbers of PIDs and platform error rate.
+
+      def calculate_cut_off(m, error_rate = 0.02)
+        n = 0
+        case error_rate
+        when 0.005...0.015
+          if m <= 10
+            n = 2
+          else
+            n = 1.09*10**-26*m**6 + 7.82*10**-22*m**5 - 1.93*10**-16*m**4 + 1.01*10**-11*m**3 - 2.31*10**-7*m**2 + 0.00645*m + 2.872
+          end
+
+        when 0...0.005
+          if m <= 10
+            n = 2
+          else
+            n = -9.59*10**-27*m**6 + 3.27*10**-21*m**5 - 3.05*10**-16*m**4 + 1.2*10**-11*m**3 - 2.19*10**-7*m**2 + 0.004044*m + 2.273
+          end
+
+        else
+          if m <= 10
+            n = 2
+          elsif m <= 8500
+            n = -1.24*10**-21*m**6 + 3.53*10**-17*m**5 - 3.90*10**-13*m**4 + 2.12*10**-9*m**3 - 6.06*10**-6*m**2 + 1.80*10**-2*m + 3.15
+          else
+            n = 0.0079 * m + 9.4869
+          end
+        end
+
+        n = n.round
+        n = 2 if n < 3
+        return n
+      end
+
+      # identify which file in the directory is R1 file, and which is R2 file based on file names
+      # input as directory (Dir object or a string of path)
+      # by default, .gz files will be unzipped.
+      # return as an hash of {r1_file: file1, r1_file: file2}
+      def r1r2(directory, unzip = true)
+        files = []
+        Dir.chdir(directory) { files = Dir.glob "*" }
+        r1_file = ""
+        r2_file = ""
+        files.each do |f|
+          tag = parser_file_name(f)[:tag]
+
+          if tag.include? "R1"
+            unzip ? r1_file = unzip_r(directory, f) : r1_file = File.join(directory, f)
+          elsif tag.include? "R2"
+            unzip ? r2_file = unzip_r(directory, f) : r2_file = File.join(directory, f)
+          end
+        end
+        return { r1_file: r1_file, r2_file: r2_file }
+      end # end of ViralSeq:TcsCore.r1r2
+
+      # sort directories containing mulitple r1 and r2 files.
+      # use the library name (first string before "_") to seperate libraries
+      # out_dir is the Dir object or string of the output directory, by default named as directory + "_sorted"
+      # return a hash as { with_both_r1_r2: [lib1, lib2, ...], missing_r1: [lib1, lib2, ...], missing_r2: [lib1, lib2, ...], error: [lib1, lib2, ...]}
+
+      def sort_by_lib(directory, out_dir = directory + "_sorted")
+        Dir.mkdir(out_dir) unless File.directory?(out_dir)
+        files = []
+        Dir.chdir(directory) {files = Dir.glob("*")}
+
+        files.each do |file|
+          path = File.join(directory,file)
+          index = file.split("_")[0]
+          index_dir = File.join(out_dir, index)
+          Dir.mkdir(index_dir) unless File.directory?(index_dir)
+          File.rename(path, File.join(index_dir, file))
+        end
+
+        return_obj = { with_both_r1_r2: [],
+                       missing_r1: [],
+                       missing_r2: [],
+                       error: []
+                      }
+
+        libs = []
+        Dir.chdir(out_dir) { libs = Dir.glob('*') }
+        libs.each do |lib|
+          file_check = ViralSeq::TcsCore.r1r2(File.join(out_dir, lib))
+          if !file_check[:r1_file].empty? and !file_check[:r2_file].empty?
+            return_obj[:with_both_r1_r2] << lib
+          elsif file_check[:r1_file].empty? and !file_check[:r2_file].empty?
+            return_obj[:missing_r1] << lib
+          elsif file_check[:r2_file].empty? and !file_check[:r1_file].empty?
+            return_obj[:missing_r2] << lib
+          else
+            return_obj[:error] << lib
+          end
+        end
+        return return_obj
+      end
+
+      # sort array of file names to determine if there is potential errors
+      # input name_array array of file names
+      # output hash { }
+      # need to change for each file name have an error code. and a bool to show if all pass
+      def validate_file_name(name_array)
+        errors = {
+                   file_type_error: [] ,
+                   missing_r1_file: [] ,
+                   missing_r2_file: [] ,
+                   extra_r1_r2_file: [],
+                   no_region_tag: [] ,
+                   multiple_region_tag: []
+                 }
+
+        passed_libs = {}
+
+        name_with_r1_r2 = []
+
+        name_array.each do |name|
+          tag = parser_file_name(name)[:tag]
+          if name !~ /\.fastq\Z|\.fastq\.gz\Z/
+            errors[:file_type_error] << name
+          elsif tag.count("R1") == 0 and tag.count("R2") == 0
+            errors[:no_region_tag] << name
+          elsif tag.count("R1") > 0 and tag.count("R2") > 0
+            errors[:multiple_region_tag] << name
+          elsif tag.count("R1") > 1 or tag.count("R2") > 1
+            errors[:multiple_region_tag] << name
+          else
+            name_with_r1_r2 << name
+          end
+        end
+
+        libs = {}
+
+        name_with_r1_r2.map do |name|
+          libname = parser_file_name(name)[:libname]
+          libs[libname] ||= []
+          libs[libname] << name
+        end
+
+        libs.each do |libname, files|
+          count_r1_file = 0
+          count_r2_file = 0
+          files.each do |name|
+            tag = parser_file_name(name)[:tag]
+            if tag.include? "R1"
+              count_r1_file += 1
+            elsif tag.include? "R2"
+              count_r2_file += 1
+            end
+          end
+
+          if count_r1_file > 1 or count_r2_file > 1
+            errors[:extra_r1_r2_file] += files
+          elsif count_r1_file.zero?
+            errors[:missing_r1_file] += files
+          elsif count_r2_file.zero?
+            errors[:missing_r2_file] += files
+          else
+            passed_libs[libname] = files
+          end
+        end
+
+        passed_names = []
+
+        passed_libs.values.each { |names| passed_names += names}
+
+        if passed_names.size < name_array.size
+          pass = false
+        else
+          pass = true
+        end
+
+        return { errors: errors, all_pass: pass, passed_names: passed_names, passed_libs: passed_libs }
+      end
+
+      # filter r1 raw sequences for non-specific primers.
+      # input r1_sh, SeqHash obj.
+      # return filtered Hash of sequence name and seq pair, in the object { r1_filtered_seq: r1_filtered_seq_pair }
+
+      def filter_r1(r1_sh, forward_primer)
+        if forward_primer.match(/(N+)(\w+)$/)
+          forward_n = $1.size
+          forward_bio_primer = $2
+        else
+          forward_n = 0
+          forward_bio_primer = forward_primer
+        end
+        forward_bio_primer_size = forward_bio_primer.size
+        forward_starting_number = forward_n + forward_bio_primer_size
+        forward_primer_ref = forward_bio_primer.nt_parser
+
+        r1_passed_seq = {}
+        r1_raw = r1_sh.dna_hash
+
+        proc_filter = proc do |name|
+          seq = r1_raw[name]
+          next unless general_filter seq
+          primer_region_seq = seq[forward_n, forward_bio_primer_size]
+          if primer_region_seq =~ forward_primer_ref
+            new_name = remove_tag name
+            r1_passed_seq[new_name] = seq
+          end
+        end
+
+        r1_raw.keys.map do |name|
+          proc_filter.call name
+        end
+
+        return { r1_passed_seq: r1_passed_seq, forward_starting_number: forward_starting_number }
+      end # end of filter_r1
+
+      # filter r2 raw sequences for non-specific primers.
+      # input r2_sh, SeqHash obj.
+      # return filtered Hash of sequence name and seq pair, as well as the length of PID.
+      def filter_r2(r2_sh, cdna_primer)
+        r2_raw = r2_sh.dna_hash
+        cdna_primer.match(/(N+)(\w+)$/)
+        pid_length = $1.size
+        cdna_bio_primer = $2
+        cdna_bio_primer_size = cdna_bio_primer.size
+        reverse_starting_number = pid_length + cdna_bio_primer_size
+        cdna_primer_ref = cdna_bio_primer.nt_parser
+        r2_passed_seq = {}
+        proc_filter = proc do |name|
+          seq = r2_raw[name]
+          next unless general_filter seq
+          primer_region_seq = seq[pid_length, cdna_bio_primer_size]
+          if primer_region_seq =~ cdna_primer_ref
+            new_name = remove_tag name
+            r2_passed_seq[new_name] = seq
+          end
+        end
+
+        r2_raw.keys.map do |name|
+          proc_filter.call name
+        end
+
+        return { r2_passed_seq: r2_passed_seq, pid_length: pid_length, reverse_starting_number: reverse_starting_number }
+      end # end of filter_r2
+
+
+
+      # puts error message in the log file handler, and abort with the same infor
+
+      def log_and_abort(log, infor)
+        log.puts Time.now.to_s + "\t" + infor
+        log.close
+        abort infor.red.bold
+      end
+
+      private
+
+      def unzip_r(indir, f)
+        r_file = File.join(indir, f)
+        if f =~ /.gz/
+          `gzip -d #{r_file}`
+          new_f = f.sub ".gz", ""
+          r_file = File.join(indir, new_f)
+        end
+        return r_file
+      end
+
+      def parser_file_name(file_name)
+        t = file_name.split(".")[0].split("_")
+        if t.size == 1
+          libname = "lib"
+          tag = [ t[0].upcase ]
+        else
+          libname = t[0]
+          tag = t[1..-1].map(&:upcase)
+        end
+        return {libname: libname, tag: tag}
+      end
+
+      def general_filter(seq)
+        if seq[1..-2] =~ /N/ # sequences with ambiguities except the 1st and last position removed
+          return false
+        elsif seq =~ /A{11}/ # a string of poly-A indicates adaptor sequence
+          return false
+        elsif seq =~ /T{11}/ # a string of poly-T indicates adaptor sequence
+          return false
+        else
+          return true
+        end
+      end
+
+      # remove region info tags from the raw MiSeq sequences.
+      def remove_tag(seq_name)
+        if seq_name =~ /\s/
+          new_tag = $`
+        else
+          new_tag = seq_name[0..-3]
+        end
+      end
+
+    end # end of class << self
+
+  end # end of TcsCore module
+
+end # end of main module