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multiPrime

multiPrime is an error-tolerant primer design tool for broad-spectrum pathogen detection. It propose a solution for the minimum degeneracy degenerate primer design with error (MD-EDPD).

1. Install

pip

pip3 install multiPrime
  • pip python >=3.9

2. Usage

$ multiPrime 
multiPrime Dprime-i input -o output
           Options: { -l [18] -n [4] -d [10] -v [1] -e [3.6] -g [0.2,0.7] -f [0.8] -c [4] -p [10] -a [4] }

Parameters:

Parameters Description
-i/--input Input file: Result of multi-alignment. (muscle, mafft or others)
-l/--plen Length of primer. Default: 18
-n/--dnum Number of degenerate. Default: 4.
-v/--variation Max mismatch number of primer. Default: 1.
-e/--entropy Entropy is actually a measure of disorder. This parameter is used to judge whether the window is conservation. Entropy of primer-length window. Default: 3.6.
-g/--gc Filter primers by GC content. Default [0.2,0.7].
-s/--size Number of degenerate. Default: 4.
-f/--fraction Filter primers by match fraction (Coverage with errors). Default: 0.8.
-c/--coordinate Mismatch index is not allowed to locate in start or stop. otherwise, it won't be regard as the mis-coverage. With this param, you can control the index of Y-distance (number=variation and position of mismatch) when calculate coverage with error.Default: 4.
-p/--proc Number of process to launch. Default: 20.
-a/--away Filter hairpin structure, which means distance of the minimal paired bases. Default: 4. Example:(number of X) AGCT[XXXX]AGCT. Primers should not have complementary sequences (no consecutive 4 bp complementarities),otherwise the primers themselves will fold into hairpin structure.
-o/--out Output file: candidate primers. e.g. [*].candidate.primers.txt.

3. Results

Three output files:

  • output:Information of primer.
  • output.gap_seq_id_json: Positions and non-contained sequences caused by errors (number of errors are greater than threshold).
  • output.non_coverage_seq_id_json: Positions and non-contained sequences.

4. test dir

multiPrime/example

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