Berkeley STATISTICS 246 - Introduction to microarray technology Extra material (54 pages)

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Introduction to microarray technology Extra material



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Introduction to microarray technology Extra material

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Pages:
54
School:
University of California, Berkeley
Course:
Statistics 246 - Statistical Genetics
Statistical Genetics Documents

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Introduction to microarray technology Extra material Lecture 17 Statistics 246 March 18 2004 1 Back to cDNA arrays the M Guide Build your own arrayer M Guide Array Maker Documentation Printing Microarrays 2 Printing Microarrays Print Head Plate Handling XYZ positioning Repeatability Accuracy Resolution Environmental Control Humidity Dust Instrument Control Sample Tracking Software 3 Ngai Lab arrayer UC Berkeley 4 Microarray Gridder 5 Slide Preparation Home Grown Protocol for preparing poly L Lysine slides for Microarrays 1 Wash 180 slides completely 2 Prepare poly lysine solution 3 Pour solution over slide 4 Rinse spin dry and store slides 5 Use slides no less than 2 and no more than 4 6 months later 6 7 Product Amplification and preparation What to Print Protocol for Amplifying Products to Print on Array All PCR reactions in 96 well format 100 ml reaction volume Perform PCR reactions in a Tetrad Machine Reactions are assayed on 96 well agarose gel Need multi channel pipetting system Also desirable to have Multimek 96 well pipetting robot 8 MJ Tetrad PCR machine 9 Protocol for preparation of Plasmid DNA from Bacterial Clones Containing Mammalian DNA 1 Inoculate a deep 96 well plate filled with IB antibiotic marker with a small amount of bacterial culture Incubate with shaking at 37 C 2 Spin down the cultures and follow the manufacturer s protocol for the QIAprep 3 Use 1 5 ul of eluted plasmid DNA as PCR template 10 Protocol for precipitation and 384 Well Arraying of PCR products 1 After running reactions on 1 agarose gel and documenting results add sodium acetate pH 5 5 and 110 ul room temp isopropanol 2 Transfer reactions to U bottom plates tape plates together 3 Spin plates at 4 500 rpm for 2 hours 4 Carefully aspirate solution 5 Add 100ul 70 EtOH Spin plates for another hour at 4 500 6 Aspirate again and let air dry or dry in a 96 well speed vac 7 Allow PCR products to resuspend in 20ul of H2O for at least 18 hours 8 Transfer products to 384 well printing plates



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