Dylan Maghini

January 12, 2016 RNAseq protocol:

   Flash freeze tissue from six snakes, two organs per snake
   RNA range: 100 ng to 10 micrograms
       --> how do we know that the correct portion of tissue was sampled?
   Convert RNA to mRNA using polyT oligonucleotides attached to beads, such that mRNA base pairs with beads
   Convert mRNA to cDNA with RT (reverse transcriptase, uses RNA as a template to make DNA) and dNTP)

   -RNA fragmentation, cDNA synthesis, and clean-up
       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
       How to prime every RNA simultaneously?