Optimization of a multiplex minisequencing protocol for population studies and medical genetics
Genet. mol. res. (Online)
;
4(2): 115-125, 30 jun. 2005. tab, ilus, graf
Article
in English
| LILACS
| ID: lil-445299
ABSTRACT
Several technologically sophisticated high-throughput techniques have been recently developed for the study of human single nucleotide polymorphisms and the diagnosis of point mutations in human diseases. However, there is also a need for simple and inexpensive techniques suitable for clinical services and small research laboratories. Minisequencing meets the latter requirements. It is simple, non-radioactive and can be easily multiplexed by adding oligonucleotide tails of increasing size to the sequencing oligonucleotide primers. To optimize the minisequencing protocol, we designed a test multiplex system capable of typing simultaneously 12 different human autosomal single nucleotide polymorphisms. We discovered that the quality of minisequencing primers and the careful selection of the tail sequences were especially critical for success. This optimized protocol permits rapid genotyping at low cost and can serve as a blueprint for the creation of multiplex minisequencing systems suitable to virtually any typing application in population studies and medical genetics.
Full text:
Available
Index:
LILACS (Americas)
Main subject:
Polymerase Chain Reaction
/
Sequence Analysis, DNA
/
Point Mutation
/
DNA Primers
/
Polymorphism, Single Nucleotide
Type of study:
Practice guideline
Limits:
Humans
Language:
English
Journal:
Genet. mol. res. (Online)
Journal subject:
Molecular Biology
/
Genetics
Year:
2005
Type:
Article
Affiliation country:
Brazil
Institution/Affiliation country:
Núcleo de Genética Médica de Minas Gerais/BR
/
Universidade Federal de Minas Gerais/BR
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