22Aug 2022

TOUCHDOWN PCR FOR INCREASED SENSITIVITY AND SPECIFICITY IN AMPLIFICATION OF MOUSE COGNATE HOMEOBOX

  • Department of Biotechnology, K L University, Guntur District, Andhra Pradesh, India.
  • Department of Bioengineering and Technology, TRC, NERECampus, Guwahati, Assam, India.
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Mammalian system have a large genome with a high level of gene sequence identity from other genomic DNA, making assessment difficult and time-consuming.Our findings describe a simple method for rapidly isolating and amplifying HoxA loci in the mouse genome using degenerate primers. For the semi degenerate primers,they were designed based on cognate gene coding regions of consensus sequences. After assembling sequences from different primer matches amplifying the same HoxA loci, the effects of the universal primer-template match on the efficiency of standard PCR amplification were investigated. Touchdown PCR increased specificity and yield in two consecutive amplifications on different gel concentrations by using high and low annealing temperatures. This method was quick, simple, and inexpensive for amplification of consensus sequences in very large gene sequences.


[Sri Sai Priya Avuthu, Bharathi Yelem, Sai Jathin Nunagonda, Sai Bhumika Reddy Battagiri, Monyya Sree Kommineni, Jythri Dandamudi, Sahithi Mulpuri and Reethika Singh Ranwas (2022); TOUCHDOWN PCR FOR INCREASED SENSITIVITY AND SPECIFICITY IN AMPLIFICATION OF MOUSE COGNATE HOMEOBOX Int. J. of Adv. Res. 10 (Aug). 699-703] (ISSN 2320-5407). www.journalijar.com


Reethika Singh Ranwas
Department of Bioengineering and Technology, TRC, NERE Campus, Guwahati, Assam, India
India

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Article DOI: 10.21474/IJAR01/15224      
DOI URL: http://dx.doi.org/10.21474/IJAR01/15224