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volume 47 issue 1 (february 2013) : 91-93

OPTIMIZATION OF SINGLE-STRAND CONFORMATION POLYMORPHISM (SSCP) ANALYSIS FOR HSP90AA1 GENE OF DEONI CATTLE IN THE PRESENCE OF POLYETHYLENE GLYCOL

S
Shahid Ahmad Shergojry*
K
K.P. Ramesha
M
M. Raies Haq
A
Ajaz. A Ganie
B
Bashir A. Ganie
P
Peerzada Altaf Q
1National Dairy Research Institute,Southern Campus, Adugodi, Bangalore-560 030, India

  • Submitted|

  • First Online |

  • doi

Cite article:- Shergojry* Ahmad Shahid, Ramesha K.P., Haq Raies M., Ganie A Ajaz., Ganie A. Bashir, Q Altaf Peerzada (2025). OPTIMIZATION OF SINGLE-STRAND CONFORMATION POLYMORPHISM (SSCP) ANALYSIS FOR HSP90AA1 GENE OF DEONI CATTLE IN THE PRESENCE OF POLYETHYLENE GLYCOL. Indian Journal of Animal Research. 47(1): 91-93. doi: .

ABSTRACT

For the first time, we report the optimization of single-strand conformation polymorphism (SSCP) analysis for HSP90AA1 gene of Deoni cattle in the presence of polyethylene glycol (PEG). The protocol developed separates single stranded conformers with a better resolution than conventional SSCP protocols and extends the applicability of SSCP analysis from 250 to as much as 999bp of DNA. PEG modulates the rate of migration of the DNA single stranded molecules in a way that provides additional means to further control resolution. It was conclude that PEG as an aided molecule of resolution along with high resolving polyacrylamide gel electrophoresis, suggests an alternative to earlier SSCP analysis procedures. Optimized protocol could be used for genotyping, detection of mutations as well as for research purposes.

REFERENCES

  1. Budowle, B., Chakraborty, R., Giusti , A.M., Eisenberg, A.J. and Allen, R. C. (1991). Analysis of the VNTR locus DIS80 by the PCR followed by high-resolution PAGE.Am. J. Hum. Genet.48:137–44.
  2. Glavac, D. and Dean, M. (1993).Optimization of the single-strand conformation polymorphism (SSCP) technique for detection of point mutations.Hum. Mutat.2: 404-414.
  3. Kapil, D., Gupta, S.C. and Neelam G. (2010).Single Strand Confirmation Polymorphism (SSCP) detection in growth differentiation factor-9 (GDF-9) gene of Karnahsheep (OvisOriantalisVignei) in Himalayan regions. I. J. Dai. Sci.63: 10-15.
  4. Kirkpatrick, B.W. (1992). Detection of a three-allel single strand conformational polymorphism (SSCP) in the fourth intron of the bovine growth harmone gene.Anim. genet.,23: 179-181.
  5. Mukhopadhyay, C.S., Gupta, A.K., Yadav, B.R. and Mohanty, T.K. (2011). Exploration of Y-specific markers to detect SNP associated with sub fertility traits in dairy bulls. Indian J. Biotech.10: 178-182.
  6. Skowach, K., Wiegand, P. and Brinkmann, B. (1992) a new allelic ladder and an improved electrophoretic separation lead to the demonstration of 28 alleles. Int. J. Legal Med. 105: 165–168.
  7. Yang, G., Hickford, J.G., Zhou, H., Fang, Q.andForrest, R.H. (2011). Extended haplotype analysis of ovine ADRB3 using polymerase chain reaction single strand conformational polymorphism on two regions of the gene. DNA Cell Biol.30:445-448.
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    volume 47 issue 1 (february 2013) : 91-93

    OPTIMIZATION OF SINGLE-STRAND CONFORMATION POLYMORPHISM (SSCP) ANALYSIS FOR HSP90AA1 GENE OF DEONI CATTLE IN THE PRESENCE OF POLYETHYLENE GLYCOL

    S
    Shahid Ahmad Shergojry*
    K
    K.P. Ramesha
    M
    M. Raies Haq
    A
    Ajaz. A Ganie
    B
    Bashir A. Ganie
    P
    Peerzada Altaf Q
    1National Dairy Research Institute,Southern Campus, Adugodi, Bangalore-560 030, India

    • Submitted|

    • First Online |

    • doi

    Cite article:- Shergojry* Ahmad Shahid, Ramesha K.P., Haq Raies M., Ganie A Ajaz., Ganie A. Bashir, Q Altaf Peerzada (2025). OPTIMIZATION OF SINGLE-STRAND CONFORMATION POLYMORPHISM (SSCP) ANALYSIS FOR HSP90AA1 GENE OF DEONI CATTLE IN THE PRESENCE OF POLYETHYLENE GLYCOL. Indian Journal of Animal Research. 47(1): 91-93. doi: .

    ABSTRACT

    For the first time, we report the optimization of single-strand conformation polymorphism (SSCP) analysis for HSP90AA1 gene of Deoni cattle in the presence of polyethylene glycol (PEG). The protocol developed separates single stranded conformers with a better resolution than conventional SSCP protocols and extends the applicability of SSCP analysis from 250 to as much as 999bp of DNA. PEG modulates the rate of migration of the DNA single stranded molecules in a way that provides additional means to further control resolution. It was conclude that PEG as an aided molecule of resolution along with high resolving polyacrylamide gel electrophoresis, suggests an alternative to earlier SSCP analysis procedures. Optimized protocol could be used for genotyping, detection of mutations as well as for research purposes.

    REFERENCES

    1. Budowle, B., Chakraborty, R., Giusti , A.M., Eisenberg, A.J. and Allen, R. C. (1991). Analysis of the VNTR locus DIS80 by the PCR followed by high-resolution PAGE.Am. J. Hum. Genet.48:137–44.
    2. Glavac, D. and Dean, M. (1993).Optimization of the single-strand conformation polymorphism (SSCP) technique for detection of point mutations.Hum. Mutat.2: 404-414.
    3. Kapil, D., Gupta, S.C. and Neelam G. (2010).Single Strand Confirmation Polymorphism (SSCP) detection in growth differentiation factor-9 (GDF-9) gene of Karnahsheep (OvisOriantalisVignei) in Himalayan regions. I. J. Dai. Sci.63: 10-15.
    4. Kirkpatrick, B.W. (1992). Detection of a three-allel single strand conformational polymorphism (SSCP) in the fourth intron of the bovine growth harmone gene.Anim. genet.,23: 179-181.
    5. Mukhopadhyay, C.S., Gupta, A.K., Yadav, B.R. and Mohanty, T.K. (2011). Exploration of Y-specific markers to detect SNP associated with sub fertility traits in dairy bulls. Indian J. Biotech.10: 178-182.
    6. Skowach, K., Wiegand, P. and Brinkmann, B. (1992) a new allelic ladder and an improved electrophoretic separation lead to the demonstration of 28 alleles. Int. J. Legal Med. 105: 165–168.
    7. Yang, G., Hickford, J.G., Zhou, H., Fang, Q.andForrest, R.H. (2011). Extended haplotype analysis of ovine ADRB3 using polymerase chain reaction single strand conformational polymorphism on two regions of the gene. DNA Cell Biol.30:445-448.
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