Introducing microarray Gene expression There are many different

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Introducing microarray

Introducing microarray

Gene expression • There are many different cell types, but every cell in our

Gene expression • There are many different cell types, but every cell in our body contains the same genetic information. • Why? Gene expression. 2

When are genes expressed? 3

When are genes expressed? 3

Measuring gene expression Different techniques are used to measure gene expression, such as: •

Measuring gene expression Different techniques are used to measure gene expression, such as: • northern blotting • reverse transcription polymerase chain reaction (RT-PCR) • microarray 4

Microarray and gene expression • Microarray technology measures the amount of m. RNA present

Microarray and gene expression • Microarray technology measures the amount of m. RNA present in many samples at once. • What genes are expressed and in what amounts? Image courtesy of Affymetrix Inc 5

Microarray and cancer Microarray is used to study cancer, enabling: • classification of disease

Microarray and cancer Microarray is used to study cancer, enabling: • classification of disease subtypes • identification of genes that may be useful targets for personalised medical treatment • selection of treatments suitable for individual patients 6

Step 1: Prepare microarray slide A microarray slide is embedded with manufactured single-stranded DNA

Step 1: Prepare microarray slide A microarray slide is embedded with manufactured single-stranded DNA segments from known genes. 7

Step 2: Collect patient samples Normal skin cells and melanoma cells are collected from

Step 2: Collect patient samples Normal skin cells and melanoma cells are collected from each patient and m. RNA isolated. 8

Step 3: Convert and label • m. RNA in patient samples is converted into

Step 3: Convert and label • m. RNA in patient samples is converted into c. DNA. • Samples are then labelled with fluorescent dye: normal skin cells with green dye, and melanoma cells with red. Once labelled, the two c. DNA samples are mixed together. 9

Step 4: Add c. DNA to microarray slide • c. DNA will hybridise (bind)

Step 4: Add c. DNA to microarray slide • c. DNA will hybridise (bind) with complementary strands embedded on the microarray slide. 10

Step 5: Visualise microarray results • Using laser microscopy the microarray slide is scanned

Step 5: Visualise microarray results • Using laser microscopy the microarray slide is scanned revealing different coloured spots on the slide: red, green, yellow and black. 11

Interpreting microarray results 12

Interpreting microarray results 12

Microarray simulation This simulation uses p. H indicators to simulate microarray technique. gene expressed

Microarray simulation This simulation uses p. H indicators to simulate microarray technique. gene expressed more in normal cells gene expressed equally in normal cells and cancer cells gene expressed more in cancer cells 13

© 2015, The University of Western Australia ast 1166 | version 1. 0 Gene

© 2015, The University of Western Australia ast 1166 | version 1. 0 Gene expression 3: Introducing microarray For conditions of use see spice. wa. edu. au/usage Developed for the Department of Education, Western Australia 14

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