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  • OTU
    replied
    Left and right are not complements of each other. They match. That's how they overlap. For denovo assembly try using programs like SOAP denovo
    http://soap.genomics.org.cn/soapdenovo.html, or SPADES de novo http://bioinf.spbau.ru/en/. Are your sequences from single cell? Or a metagenome?

    Leave a comment:


  • mido1951
    replied
    no. i have illumina paired end reads. and I want to denovo assembly without a reference genome.
    how the right portion of a reads overlaps with the left portion of another reads? (it was not the same sense od read)
    And have you an example

    Leave a comment:


  • OTU
    replied
    Assembly algorithm will depend whether you have a reference genome or not. Is it a metagenome sequencing?

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  • mido1951
    replied
    how make an assembly with reads (paired end: two files) .
    Is it that you can make an example please?
    thanks

    Leave a comment:


  • OTU
    replied
    No. If you have a fragment of DNA: ATCGTTGAGCAGACT,
    your R1: TAGCAA
    and R2: GTCTGA

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  • mido1951
    replied
    for example:
    we have a fragment of DNA: ATCGTTGAGCAGACT
    we will sequence this fragment and for example after the paired end sequencing reads we have R1 and R2 with a length 6. Thus:
    R1 = ATCGTT
    R2 = AGTCTG (reverse complement)
    so after sequencing wa have R1..... R2 (the middle part is unknown).
    that's the paired end reads.
    or not?

    Leave a comment:


  • OTU
    replied
    Oh, that's just an example of what paired-end pairing looks like.

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  • mido1951
    replied
    how you got these reads !! ??

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  • OTU
    replied
    Then you should consider that overlap is something like:
    R1-ATTGCTGTG
    -----------ACACTGAAAAGT-R2

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  • mido1951
    replied
    I speak of an example.
    it is assumed that it is an overlap.
    I want to create an assembly tool but first I need to know how to detect overlap between the paired ends (from two files). and make assembly with paired end.

    Leave a comment:


  • OTU
    replied
    Don't see why
    "F1.fq:
    S1: R1=ATCGTTGAG
    S2: R1=TGAGCAGAC " would overlap. They only match at 3 bp. Assemblers won't combine them.

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  • mido1951
    replied
    for example:
    we have the sequence: S1: ATCGTTGAGCAGACT and the sequence S2: TGAGCAGACTTAAGTAGTTTT .
    and for example, was the first sequenced reads from S1: R1 = ATCGTTGAG
    R2 = AGTCTGCTC (reverse complement from the right)
    and from the second sequence: R1: TGAGCAGAC
    R2: AAAACTACT (reverse complement from the right)
    So we have the two files paired end:
    F1.fq:
    S1: R1=ATCGTTGAG
    S2: R1=TGAGCAGAC
    F2.fq:
    S1: R2=AGTCTGCTC
    S2: R2=AAAACTACT

    in the assembly here there is an overlap between R1(S1) and R1(S2).
    in assembly, we can have overlap between R1 and R2 from two differents sequence??
    Last edited by mido1951; 10-22-2015, 01:55 PM.

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  • GenoMax
    replied
    Originally posted by mido1951 View Post
    I have llumina paired end data.
    I want to make an assembly of these data.
    But the problem I do not understand the two F1.fq file and F2.fq.
    Is that reads and reads of F1.fq F2.fq are complementary or not?
    for the assembly do I have to overlap F1.fq or I have to overlap and F1.fq F2.fq?
    thanky
    Cross-posted: https://www.biostars.org/p/162806/

    @mido1951: See this page for a simple explanation of "shotgun sequencing": https://en.wikipedia.org/wiki/Shotgun_sequencing In the past people used sanger sequencing for this, which has now been replaced with NGS.

    R1/R2 are merely sequences from the two ends of a fragment. They do not need to be complementary (in fact in most cases they will not be). You do not need to worry about R1/R2 reads individually but use them as a set for assembly.

    Leave a comment:


  • mido1951
    replied
    I have llumina paired end data.
    I want to make an assembly of these data.
    But the problem I do not understand the two F1.fq file and F2.fq.
    Is that reads and reads of F1.fq F2.fq are complementary or not?
    for the assembly do I have to overlap F1.fq or I have to overlap and F1.fq F2.fq?
    thanky

    Leave a comment:


  • OTU
    replied
    What is your data on? Metagenome, single genome?
    What sequencing platform did you use? What is the processing computer power that you can use?

    Leave a comment:

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