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| − | '''January 12, 2016''' | + | '''Class Notes''' |
| − | RNAseq protocol:
| + | [[Group 1 intestines]] |
| − | Flash freeze tissue from six snakes, two organs per snake
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| − | RNA range: 100 ng to 10 micrograms
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| − | --> how do we know that the correct portion of tissue was sampled?
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| − | Convert RNA to mRNA using polyT oligonucleotides attached to beads, such that mRNA base pairs with beads
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| − | Convert mRNA to cDNA with RT (reverse transcriptase, uses RNA as a template to make DNA) and dNTP)
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| | | | |
| − | -RNA fragmentation, cDNA synthesis, and clean-up
| + | [[Literature summary]] |
| − | high throughput sequencing has only ~75bp of accurate reads
| + | |
| − | fragmentation takes one long mRNA and cuts it into similarly sized fragments so that there are more edges to read from
| + | [[DM Notes 1.12.16]] |
| − | How to prime every RNA simultaneously? Generate every possible hexamer for the primers (4^6 different combinations)
| + | |
| − | Product:
| + | [[DM Notes 1.14.16]] |
| − | Look at images in pptx
| + | |
| − | Second band of images: gel purified RNA
| + | [[DM Notes 1.21.16]] |
| − | Third band of images: reamplified (considered good cDNA library for RNAseq)
| + | |
| | + | [[DM Notes 1.26.16]] |
| | + | |
| | + | [[DM Notes 1.28.16]] |
| | + | |
| | + | [[DM Notes 2.02.16]] |
| | + | |
| | + | [[DM Notes 2.04.16]] |
| | + | |
| | + | [[DM Notes 2.09.16]] |
| | + | |
| | + | [[DM Notes 2.11.16]] |
| | + | |
| | + | [[DM Notes 2.16.16]] |
| | + | |
| | + | [[DM Notes 2.18.16]] |
| | + | |
| | + | [[DM Notes 2.23.16]] |
| | + | |
| | + | [[DM Notes 2.25.16]] |
| | + | |
| | + | [[DM Notes 3.8.16]] |
| | + | |
| | + | [[DM Notes 3.10.16]] |
| | + | |
| | + | [[DM Notes 3.17.16]] |
| | + | |
| | + | [[DM Notes 3.22.16]] |
| | + | |
| | + | [[DM Notes 3.24.16]] |
| | + | |
| | + | [[DM Notes 3.31.16]] |
| | + | |
| | + | [[DM Notes 4.5.16]] |
| | + | |
| | + | [[DM Notes 4.7.16]] |
Latest revision as of 17:04, 7 April 2016
Class Notes
Group 1 intestines
Literature summary
DM Notes 1.12.16
DM Notes 1.14.16
DM Notes 1.21.16
DM Notes 1.26.16
DM Notes 1.28.16
DM Notes 2.02.16
DM Notes 2.04.16
DM Notes 2.09.16
DM Notes 2.11.16
DM Notes 2.16.16
DM Notes 2.18.16
DM Notes 2.23.16
DM Notes 2.25.16
DM Notes 3.8.16
DM Notes 3.10.16
DM Notes 3.17.16
DM Notes 3.22.16
DM Notes 3.24.16
DM Notes 3.31.16
DM Notes 4.5.16
DM Notes 4.7.16
