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    PRSV

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    Balaji Cg
    Individual leaves from papaya plants were collected for RNA extraction. RNA was extracted from young leaves using a genei pure RNA column method. The extracted RNA was converted to cDNA using a First stand cDNA Synthesize Kit. PCR was performed using primers specific for papaya virus to amplify a region of around 1.1 kb. The PCR conditions included an initial denaturation at 94°C followed by 35 cycles of denaturation, annealing and extension before a final elongation step of 30 minutes at 72°C. Gel electrophoresis showed an amplified product of approximately 1.1 kb, indicating the presence of papaya virus in the infected plant samples.

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    PRSV

    Uploaded by

    Balaji Cg
    14 views2 pages
    Individual leaves from papaya plants were collected for RNA extraction. RNA was extracted from young leaves using a genei pure RNA column method. The extracted RNA was converted to cDNA using a First stand cDNA Synthesize Kit. PCR was performed using primers specific for papaya virus to amplify a region of around 1.1 kb. The PCR conditions included an initial denaturation at 94°C followed by 35 cycles of denaturation, annealing and extension before a final elongation step of 30 minutes at 72°C. Gel electrophoresis showed an amplified product of approximately 1.1 kb, indicating the presence of papaya virus in the infected plant samples.

    Original Description:

    To know about papaya ringspot virus detection through molecular studies

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    Individual leaves from papaya plants were collected for RNA extraction. RNA was extracted from young leaves using a genei pure RNA column method. The extracted RNA was converted to cDNA using a First stand cDNA Synthesize Kit. PCR was performed using primers specific for papaya virus to amplify a region of around 1.1 kb. The PCR conditions included an initial denaturation at 94°C followed by 35 cycles of denaturation, annealing and extension before a final elongation step of 30 minutes at 72°C. Gel electrophoresis showed an amplified product of approximately 1.1 kb, indicating the presence of papaya virus in the infected plant samples.

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    © All Rights Reserved
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    14 views2 pages

    PRSV

    Uploaded by

    Balaji Cg
    Individual leaves from papaya plants were collected for RNA extraction. RNA was extracted from young leaves using a genei pure RNA column method. The extracted RNA was converted to cDNA using a First stand cDNA Synthesize Kit. PCR was performed using primers specific for papaya virus to amplify a region of around 1.1 kb. The PCR conditions included an initial denaturation at 94°C followed by 35 cycles of denaturation, annealing and extension before a final elongation step of 30 minutes at 72°C. Gel electrophoresis showed an amplified product of approximately 1.1 kb, indicating the presence of papaya virus in the infected plant samples.

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    Papaya

    (Carica papaya)
    RNA Isolation:
    o Individual leaves of plants were collected for RNA
    micro-extraction. RNA was extracted from young leaf
    by genei pure RNA column method.
    cDNA conversion
    The conversion of cDNA form total RNA with the use of
    First stand cDNA Synthesize Kit (Thermo scientific).
    PCR Reaction
    The PCR reactions were performed in a total volume of 20 L
    10L PCR mastermix (pcr Buffer MgCl2 dNTP Taq Polymerase)
    2l from each Primer

    Forward primer-PNIBF1/F -GGBAAYAATAGTGGNCAACC


    Reverse primer-PCPR1/R - GGGGAGGTGCCGTTCTCDATRCACCA
    2L DNA (2050 ng;)

    4L ddH2O.

    PCR Cycle

    The PCR was carried out for


    Initial Denaturation2 Minutes/94C, followed by 35 cycles of
    Denaturation : 1Minute/ 94C;

    Annealing

    : 1minute/55C;

    Extension

    : 2 min/72C.

    S S M

    The final elongation reaction was 30 minutes/72C.

    1.1kb
    o Amplification of around 1.1 kb in poty virus
    primers in papaya virus infected samples

    M 1kb ladder S-Sample DNA

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