Protection Chemistry and Amidites,Permanent Protection,Hydroxyl Group,Heterocyclic Base,Phosphate Nucleotides

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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

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Protection Chemistry and Amidites
The methods of gene synthesis make use of protection chemistry to minimize undesired side reactions during the synthesis. Two different types of protecting groups are needed: (i) permanent protection is needed for the primary amino groups of all heterocyclic bases and for the hydroxyl groups of the phosphate of all nucleotides; (ii) temporary protection is needed for 5' hydroxyl group of the incoming nucleotide. No other functional groups need to be protected since they do not react under the conditions used for gene synthesis. A fully protected nucleotide is called an amidite.

A, C, A,C G and T amidites are needed in all synthesizers. However β-cyanoethyl phosphoramidites using the PAC protection scheme (see later for PAC) are used in Gene Assembler Synthesizers.

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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