LECTURE 16 Flashcards
what are microarrays?
shows which genes are being actively transcribed in a sample from a cell
what can direct sequencing substitute for?
hybridisation
gives better resolution and accuracy
what is high throughput sequencing?
Rapid DNA sequencing on a micro scale in which many fragments of DNA are sequenced in parallel.
what is Illumina sequencing?
fragments of DNA bound to solid surface
How is mRNA separated from the rest of the RNA in a cell?
poly T beads that attach to poly A Tails
how is RNA sequenced?
- mRNA converted to cDNA
- cDNA used for sequencing library generation
how does RNA sequencing work?
- poly a selection
- cDNA synthesis from first and second strand
- end repair, phosphorylation and A tailings
- adapter ligation, per amplification anf sequencing
what is ChIP sequencing?
Chromatin immunoprecipitation followed by DNA sequencing
-crosslinks proteins to DNA
-isolates DNA and shears
-immunoprecipitate protein of interest
-reverse cross linking
-purify DNA
-sequence
what is ATAC sequencing?
Assay for Transposase-Accessible Chromatin
what does ATAC sequencing rely on ?
transposes Tn5
how does ATAC sequencing work?
adapted ligated fragments isolated, amplified and sequenced
what is bisulphite sequencing?
Bisulphite treatment is used to determine the methylation state of DNA
Methylated cytosine protected from deamination
Unmethylated cytosine converted to thymine (via uracil)
what does BS-seq allow identification of?
hypo and hyper methylated regions
what is Reduced Representation BS-Seq (RRBS)?
-utilises Mspl restriction enzyme to enrich for CpGs
