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    Diseño Oligonucleoticos

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    55565556
    This document discusses designing oligonucleotides for a DNA sequence from E. coli O157:H7. A 20 base pair fragment is selected from the forward strand as an oligonucleotide primer. Its melting temperature is calculated to be 64°C based on its GC content. The reverse complementary sequence is calculated to have a melting temperature of 58°C. To equalize the melting temperatures, one base may need to be added or removed from one of the oligonucleotides.

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    Diseño Oligonucleoticos

    Uploaded by

    55565556
    20 views1 page
    This document discusses designing oligonucleotides for a DNA sequence from E. coli O157:H7. A 20 base pair fragment is selected from the forward strand as an oligonucleotide primer. Its melting temperature is calculated to be 64°C based on its GC content. The reverse complementary sequence is calculated to have a melting temperature of 58°C. To equalize the melting temperatures, one base may need to be added or removed from one of the oligonucleotides.

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    Diseño oligonucleoticos

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    This document discusses designing oligonucleotides for a DNA sequence from E. coli O157:H7. A 20 base pair fragment is selected from the forward strand as an oligonucleotide primer. Its melting temperature is calculated to be 64°C based on its GC content. The reverse complementary sequence is calculated to have a melting temperature of 58°C. To equalize the melting temperatures, one base may need to be added or removed from one of the oligonucleotides.

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    Download as DOC, PDF, TXT or read online from Scribd
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    Download as doc, pdf, or txt
    20 views1 page

    Diseño Oligonucleoticos

    Uploaded by

    55565556
    This document discusses designing oligonucleotides for a DNA sequence from E. coli O157:H7. A 20 base pair fragment is selected from the forward strand as an oligonucleotide primer. Its melting temperature is calculated to be 64°C based on its GC content. The reverse complementary sequence is calculated to have a melting temperature of 58°C. To equalize the melting temperatures, one base may need to be added or removed from one of the oligonucleotides.

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    Diseño oligonucleoticos

    >gi|16445223:4019893-4021734 Escherichia coli O157:H7 EDL933, complete


    genome
    ATGGAGCAAAACCCGCAGTCACAGCTGAAACTTCTTGTCACCCGTGGTAAGGAGCAAGGCTATCTGACCT
    ATGCCGAGGTCAATGACCATCTGCCGGAAGATATCGTCGATTCAGATCAGATCGAAGACATCATCCAAAT
    GATCAACGACATGGGCATTCAGGTGATGGAAGAAGCACCGGATGCCGATGATCTGATGCTGGCTGAAAAC
    ACCGCGGACGAAGATGCTGCCGAAGCCGCCGCGCAGGTGCTTTCCAGCGTGGAATCTGAAATCGGGCGCA
    CGACTGACCCGGTACGCATGTACATGCGTGAAATGGGCACCGTTGAACTGTTGACCCGCGAAGGCGAAAT
    TGACATCGCTAAGCGTATTGAAGACGGGATCAACCAGGTTCAATGCTCCGTTGCTGAATATCCGGAAGCG
    ATCACCTATCTGCTGGAACAGTACGATCGTGTTGAAGCAGAAGAAGCGCGTCTGTCCGATCTGATCACCG
    GCTTTGTTGACCCGAACGCAGAAGAAGATCTGGCACCTACCGCCACTCACGTCGGTTCTGAGCTTTCCCA
    GGAAGATCTGGACGATGACGAAGATGAAGACGAAGAAGATGGCGATGACGACAGCGCCGATGATGACAAC
    AGCATCGACCCGGAACTGGCTCGCGAAAAATTTGCGGAACTGCGCGCTCAGTACGTTGTAACGCGTGACA
    CCATCAAAGCGAAAGGTCGCAGTCACGCTGCCGCTCAGGAAGAGATCCTGAAACTGTCTGAAGTATTCAA
    ACAGTTCCGCCTGGTGCCGAAGCAGTTTGACTACCTGGTCAACAGTATGCGCGTCATGATGGACCGCGTT
    CGTACGCAAGAACGTCTGATCATGAAGCTCTGCGTTGAGCAGTGCAAAATGCCGAAGAAAAACTTCATTA
    CTCTGTTTACCGGCAACGAAACCAGCGATACCTGGTTCAACGCGGCAATTGCGATGAACAAGCCGTGGTC
    GGAAAAACTGCACGATGTCTCTGAAGAAGTGCATCGCGCCCTGCAGAAACTGCAGCAGATTGAAGAAGAA
    ACCGGCCTGACCATCGAGCAGGTTAAAGATATCAACCGTCGTATGTCCATCGGTGAAGCGAAAGCCCGCC
    GTGCGAAGAAAGAGATGGTTGAAGCGAACTTACGTCTGGTTATTTCTATCGCCAAGAAATACACCAACCG
    TGGCTTGCAGTTCCTTGACCTGATTCAGGAAGGCAACATCGGTCTGATGAAAGCGGTTGATAAATTCGAA
    TACCGCCGTGGTTACAAGTTCTCCACCTACGCAACCTGGTGGATCCGTCAGGCGATCACCCGCTCTATCG
    CGGATCAGGCGCGCACCATCCGTATTCCGGTGCATATGATTGAGACTATCAACAAACTCAACCGTATTTC
    TCGCCAGATGCTGCAAGAGATGGGCCGTGAACCGACGCCGGAAGAACTGGCTGAACGTATGCTGATGCCG
    GAAGACAAGATCCGCAAAGTGCTGAAGATCGCCAAAGAGCCAATCTCCATGGAAACGCCGATCGGTGATG
    ATGAAGATTCGCATCTGGGGGATTTCATCGAGGATACCACCCTCGAGCTGCCGCTGGATTCTGCGACCAC
    CGAAAGCCTGCGTGCGGCAACACACGACGTGCTGGCTGGCCTGACCGCGCGTGAAGCGAAAGTTCTGCGT
    ATGCGTTTCGGTATCGATATGAACACCGATCACACGCTGGAAGAAGTGGGTAAACAGTTCGACGTTACCC
    GTGAACGTATCCGTCAGATCGAAGCGAAGGCGCTGCGCAAACTGCGTCACCCGAGCCGTTCTGAAGTGCT
    GCGTAGCTTCCTGGACGATTAA

    20 bases por estabilidad

    Oligonucleotido directo 5º---3ª

    Oligonucleotido forward: ATGGAGCAAAACCCGCAGTC

    11 ( g/c) composiciòn 9x2 = 18


    18x4 = 64ºC : TM MELTING

    Oligonucleotido reverse 5º---3º (complementario)

    GCGTAGCTTCCTGGACGATTAA -------
    !!!!!!!!!!!!!!!!!!!!!!
    CGCATCGAAGGACCTGCTAATT -------EL REVERSO QUE SE PONE COMO
    OLIGONUCLEOTIDO

    11X2=22 TTAATCGTCCAGGAAGCTAC---- EL QUE SE PONE COMO


    OLIGONUCLEOTIDO

    9X4=36 = TM: 58ºC

    Como tienen TM distinto se puede agregar o quitar un base en el


    oligonucleotido para igualar TMs pero no en 5º a menos que se quiera
    marcar.

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