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1

DNA polymorphism in locus D1S80 in Poland. DNA profiling and detection of new alleles by heteroduplex formation between alleles of the same size

100%
Kwiatkowska J., Dziechciowska K., Lisiecka D., Slomski R.
Journal of Applied Genetics
|
1997
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vol. 38
|
issue 3
335-341
EN
We have analysed allele distribution at the highly polymorphic variable number of tandem repeats (VNTR) locus D1S80 (pMCT118) in the Polish population using the polymerase chain reaction (PCR) technique. Characteristics of the D1S80 locus makes it a very useful marker for population genetic research, genetic linkage studies and forensic identification of individuals. During our routine application of the D1S80 marker to paternity testing in several cases of homozygosity detected by polyacrylamide gel electrophoresis, heteroduplex formation for alleles 18 and 24 was also observed. Direct sequencing of PCR products revealed that alleles 18 and 24 of locus D1S80 actually represent a mixture composed of different sequences. Our observations indicate that identification of some 18 and 24 VNTR alleles based only on size estimated in electrophoretic analyses could lead to errors in paternity testing and DNA profiling.
2

Polymerase chain reaction-heteroduplex (PCR-HD) polymorphism within the bovine STAT5A gene

88%
Flisikowski K., Zwierzchowski L.
Journal of Applied Genetics
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2003
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vol. 44
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issue 2
185-189
EN
Signal transducers and activators of transcription (STATs) are transcription factors mediating signals of various hormones and cytokines. STAT5A, previously known as the mammary gland factor (MGF), mediates the action of prolactin on milk protein gene expression in mammary epithelial cells. We used polymerase chain reaction-heteroduplex (PCR-HD) and sequencing methods for detection of nucleotide sequence polymorphism in intron 15 of the bovine STAT5A gene. A 281-bp gene fragment, from nt 12525 to nt 12806 (GenBank AJ 237937), was amplified with PCR, denatured and subjected to polyacrylamide gel electrophoresis to detect PCR-HD polymorphism. Three genotypes and two alleles were identified. DNA samples derived from homozygotes AA and BB were sequenced. A trinucleotide CCT deletion was found in the variant B of the STAT5A gene at position 12549. In a group of 72 beef bulls of various breeds and 49 Friesian bulls genotyped by PCR-HD mostly the AA genotype was found (from 83 to 61% depending on breed). The frequency of allele A varied between 0.91 and 0.77. Animals of genotype BB were found in Charolaise and Limousine breeds only.
3

DNA speed reading course ? high-throughput DNA sequencing technologies

88%
Kotowska M., Zakrzewska-Czerwinska M.
Biotechnologia
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2010
|
issue 4
24-38
EN
Development of high-throughput DNA sequencing technologies that omit time consuming and labour intensive cloning steps have opened unprecedented possibilities in life sciences. Massive scale generation of raw sequences requires constant improvement of computational methods of data analysis. New disciplines of genomics, metagenomics and transcriptomics have emerged which revolutionize experimental approach to different fields of biology. Both basic studies, such as species evolution or microbial ecology, and applied sciences of biotechnology and medicine greatly from the new tools available. In this article next-generation DNA sequencing technologies are reviewed. Information on data analysis and applications is also provided.
4

Genome sequencing and progress in biotechnology

75%
Przybecki Z., Pawelkowicz M. E., Woycicki R.
Biotechnologia
|
2010
|
issue 4
9-23
EN
As a result of the Human Genome Project and announcement of the project: Archon X Prize for Genomics 'genome for 1000$', the sequencing technology made huge progress. The purpose was to develop radically new technology that would dramatically reduce the time and cost of sequencing and resequencing eukaryotic genomes (or large region), and transcriptomes. This progress has influenced many areas also widely understood biotechnology. Owing to comparative genomic and sequenced genomes, new opportunities have occurred such as acceleration of genomic polymorphism identification, genes and genome structure prediction. It has also stimulated the development of assignation of functional effect of SNP, CNP or INDEL polymorphism and enabled functional and structural annotation of genome, establishment of a net of genome interaction (netoms) and netom's controlling. Genetic and physical mapping of genome elements methods undergo considerable alterations, thus they become critical in association methodology of genome and their function. The analysis of metabolic network and its elements is basic in planning activities in new biotechnology. Here we present the achievements in cucumber which were possible due to sequencing the genome of this species by our team.
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