DNA Sequencing by Hybridization Using Microarrays on DNA Chips,Single Nucleotide,Immobilizing,Parallel Synthesis

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Isolation, Sequencing and Synthesis of Genes Isolation, Sequencing and Synthesis of Genes - Introduction Isolation of Genes Isolation of Genes Coding for Specific Proteins Isolation of Genes with Known or Unknown Products having Tissue Specific Expression Isolation of Genes with Known or Unknown Products Using DNA or RNA Probes Use of Heterologus Probes Use of Synthetic Probes Isolation of Genes Coding Use of Transposable Elements Transposon Tagging T-DNA-Insertion Mutagenesis for Isolation of Plant Genes Promoter Enhancer and Gene Trap for Isolation of Genes Differential Screeing and Differential Display Technique for Isolation of Genes Subtractive Hybridizatioon for Gene Isolation Map Based Cloning for Gene Isolation Isolation of Novel Genes by Asis Data at JNU New Delhi

Home >> Biotechnology and Genomics >> Isolation, Sequencing and Synthesis of Genes >> DNA Sequencing by Hybridization Using Microarrays on DNA Chips

Your Ad Here	DNA Sequencing by Hybridization Using Microarrays on DNA Chips The main objective of several genome projects is the large scale sequence analysis of genomes and its application to individual case studies in medicine and research. Sequencing by hybridization for detection of single nucleotide polymorphisms (SNPs) using DNA microchips was developed to meet this objective. A sequencing microchip is developed either by parallel synthesis of a complete set of 8-mers (i.e. 48 = 65536) on a solid support (consult Chapter 5 for details). The dimension of the microchip can be further reduced by using printing technology and robotics. The technique involves hybridization of a short DNA fragment of unknown sequence with a set of short oligonucleotides, particularly 8-mers of known sequences.
Identification and analysis of a set of overlapping oligomers that form perfect duplexes with the DNA of interest permits reconstruction of the target DNA sequence. For example, if we wish to determine the sequence of a 17 base target DNA fragment, and hybridize it with complementary, overlapping 8-mers to form perfect duplexes, the overlap of seven or fewer adjacent 8-mers showing perfect match enables reconstruction of the complementary DNA sequences of the fragment. Oligonucleotides shorter or longer than the 8-mers can also be used for sequencing, but while the use of shorter oligos leads to decrease in stability of the duplexes formed, the use of longer oligos leads to loss of discrimination between perfect base pair matches and mismatches.
Therefore, the sequencing microchip is mainly composed of 8-mers. The hybridization is generally carried out at lower temperature, so that discrimination between perfect and imperfect duplexes can be achieved. The limitation of the above technique is that the fragment to be sequenced should be small in size, although it should still be longer than n-1, where n is the length of the oligonucleotides immobilized on microchip. Analysis of long DNA fragments may be facilitated by fragmentation of the target DNA into smaller pieces prior to hybridization experiments. However, the method cannot be applied to repeat DNAs such as microsatellites or minisatellites.	Your Ad Here

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Sequencing of a Gene or a DNA Fragment Maxam and Gilbert's Chemical Degradation Method Sanger's Dideoxynucleotide Synthetic Method Automatic DNA Sequencers Slab Gel Versus Capillary Sequencers Slab Gel Sequencing Systems Capillary Gel Sequencing Systems Direct DNA Sequencing Using PCR Ligation Mediated PCR LMPCR DNA Sequencing Through Transcription DNA Sequencing by Hybridization Using Microarrays on DNA Chips DNA Sequencing by DE-MALDI-TOF Mass Spectrometry Synthesis of Genes Synthesis of Gene for Yeast Alanyl tRNA Synthesis of Oligonucleotides Synthesis of Three Duplex Fragments Synthesis of Gene from Three Duplex Fragments Synthesis of Gene for a True Precursor tRNA Total Synthesis of a Human Leukocyte Interferon Gene Gene Synthesis Machines Protection Chemistry and Amidites Permanent Protecting Groups Temporary Protecting Groups Synthesis of Gene Using PCR Synthesis of Genes From mRNA

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