TY - CHAP
T1 - The Significance of PCR Primer Design in Genetic Diversity Studies
T2 - Exemplified by Recent Research into the Genetic Structure of Marine Species
AU - Delghandi, Madjid
AU - Delghandi, Marit Pedersen
AU - Goddard, Stephen
N1 - Publisher Copyright:
© 2022, Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2021/11/14
Y1 - 2021/11/14
N2 - Genetic markers are widely applied in the study of genetic diversity for many species. The approach incorporates a Polymerase Chain Reaction (PCR) amplification of targeted sequences in the genome. Crucial for the overall success of a PCR experiment is the careful design of synthetic oligonucleotide primers. Ideally designed primer pairs will ensure the efficiency and specificity of the amplification reaction, resulting in a high yield of the desired amplicon. Important criteria such as primer-sequence, −length, and -melting temperature (Tm) are fundamental for the selection of primers and amplification of targeted nucleotide sequences from a DNA template. There are many computational tools available to assist with critical bioinformatics issues related to primer design. These resources allow the user to define parameters and criteria that need to be taken into account when designing primers. Following the initial in silico selection, a primer pair should be further tested in vivo for their amplification efficiency and robustness. Using examples taken from genetic diversity studies in a marine crustacean, this chapter provides outlines for the application of PCR technology and discusses details for the design of primers for the development and characterization of microsatellite and SNP-markers.
AB - Genetic markers are widely applied in the study of genetic diversity for many species. The approach incorporates a Polymerase Chain Reaction (PCR) amplification of targeted sequences in the genome. Crucial for the overall success of a PCR experiment is the careful design of synthetic oligonucleotide primers. Ideally designed primer pairs will ensure the efficiency and specificity of the amplification reaction, resulting in a high yield of the desired amplicon. Important criteria such as primer-sequence, −length, and -melting temperature (Tm) are fundamental for the selection of primers and amplification of targeted nucleotide sequences from a DNA template. There are many computational tools available to assist with critical bioinformatics issues related to primer design. These resources allow the user to define parameters and criteria that need to be taken into account when designing primers. Following the initial in silico selection, a primer pair should be further tested in vivo for their amplification efficiency and robustness. Using examples taken from genetic diversity studies in a marine crustacean, this chapter provides outlines for the application of PCR technology and discusses details for the design of primers for the development and characterization of microsatellite and SNP-markers.
KW - Genetic markers
KW - Marine species
KW - Microsatellite markers
KW - Molecular tools
KW - Oligonucleotide design
KW - Polymerase Chain Reaction
KW - Population genetics studies
KW - SNP markers
KW - Genetic Variation
KW - Computational Biology
KW - DNA Primers/genetics
KW - Genome
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UR - https://www.mendeley.com/catalogue/9ef13a76-b1ba-3dae-b27b-9f945435d0ba/
U2 - 10.1007/978-1-0716-1799-1_1
DO - 10.1007/978-1-0716-1799-1_1
M3 - Chapter
C2 - 34773611
AN - SCOPUS:85119355299
VL - 2392
T3 - Methods in molecular biology (Clifton, N.J.)
SP - 3
EP - 15
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -