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  • titaji
    Junior Member
    • Aug 2019
    • 2

    #1

    Primer3 issue

    Dear all

    I'm new here and I have a problem with Primer3 program.

    I'm using MISA and Primer3 in pipeline. After I got *.misa file, I put this syntax.

    Code:
    p3_in.pl FASTA.fa.misa
    Then I got empty file of *.p3in. Is it normal?

    By the way, I still tried to run further syntax.

    Code:
    primer3_core < FASTA.fa.p3in > FASTA.fa.p3out
    This one showed me the description and usage of Primer3.

    I put further syntax which is...

    Code:
    p3_out.pl FASTA.fa.p3out FASTA.fa.misa
    It showed the error message like this.

    Use of uninitialized value $count in concatenation (.) or string at C:\Users\admin\Documents\Work\NGS_results\MISA\p3_out.pl line 76.

    Primer modelling was successful for sequences.
    Use of uninitialized value $count_failed in concatenation (.) or string at C:\Users\admin\Documents\Work\NGS_results\MISA\p3_out.pl line 77.
    Primer modelling failed for sequences.
    Could anyone please help me to solve this issue?

    Thank you,
    titaji

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