Biotechnology Chapter 4 Day 3 Questions
- Most of the PCR protocols we’ve learned about using two oligonucleotides and overlap PCR required three oligonucleotides. Why does a sequencing PCR require one single primer?
- Would you expect a Sanger sequencing reaction to have equal numbers of deoxynucleotides and dideoxynucleotides? If not, which type of nucleotide should be more abundant?
- Bill Clinton threw a party in the Rose Garden in the summer of 2000 to celebrate the publication of a draft of the human genome. Two groups were given credit for this accomplishment. What was the difference in the approach taken by these groups to accomplish this feat?
- Rooms full of sequencing machines containing 384 acrylamide gels each were used to sequence the human genome. Contrast this with “massively parallel sequencing” of next-generation sequencing technologies.
- If more than one sample is included in a NGS “run”, how do you know which sample a sequencing signal came from?
- Compare how PCR reactions end up spaced apart on a 2-dimensional (planar) surface in 454 and Illumina sequencing approaches.
- What is the advantage of paired-end reads over single-end reads?
- What is read depth?
- What caveats come with the short read lengths in NGS compared to the longer read lengths in Sanger sequencing?