(精选)Transcriptomics(Gene Expression Profiling)教学课件.pptVIP

Definitions (transcriptome, transcriptomics, EST, cDNA, RT-PCR) Transcriptomics – what is it good for? Overview about transcript regulation (eukaryotic gene structure, transcription factors, micro-RNAs) Methods for measuring gene expression (single genes with qPCR, RT-PCR, Northern blotting, multiple genes with cDNA- or oligonucleotide arrays, cDNA-AFLP, DD-RTPCR, SAGE, MPSS);Part 1 – Transcriptomics (Gene Expression Profiling);Genome = entire DNA sequence of an organism Transcriptome = percentage of the genetic code that is transcribed into RNA molecules (depends on development, environment, time of the day, tissue) = collection of all gene transcripts present in a given cell/tissue at a given time (“snapshot”) Transcriptomics = global analysis of gene expression = genome-wide expression profiling cDNA = complementary DNA synthesized from mature mRNA by the enzyme reverse transcriptase cDNA library = a population of bacterial transformants or phage lysates in which each mRNA isolated form an organism or tissue is represented as its cDNA insertion in a plasmid or a phage vector;RT-PCR = a one or two-step process for converting mRNA to cDNA and the subsequent amplifcation of the reversely-transcribed DNA by PCR EST (Expressed Sequence Tag) = small pieces of DNA (200 - 500 bp) generated by sequencing either one or both ends (5’EST and 3’EST) of an expressed gene (cDNA) = can be used to identify unknown genes and to map their positions within a genome Hybridization = based on complementary molecules, sequences that are able to base-pair with one another. Adenine is the complement of thymine, guanine is the complement of cytosine. Therefore, the complementary sequence to G-T-C-C-T-A will be C-A-G-G-A-T. When two complementary sequences find each other, they will lock together, or hybridize. ;PCR (Polymerase chain reaction) = technique that results in the exponential amplification of almost any region of a selected DNA molecule;DNA; get an understanding of genes