Lecture 21 Basics: Biotechnology
21.1 Biotech Classics: Restriction enzymes and gel electrophoresis
Different restriction enzymes cut the DNA in different locations.
A. No restriction enzymes
B.
C.
21. 2 Biotech Classics: Sanger DNA sequencing
Ingredients:
Lecture 21 Page 1
Steps:
1.
2.
3.
Lecture 21 Page 2
21.3: Modern Biotechnology: Genomics, transcriptomics, proteomics
Genomics:
Transcriptomics
Proteomics:
21.4 Genomics: Next-Generation DNA Sequencing
(Watch “sales pitch” video http://youtu.be/v8p4ph2MAvI)
What are the two “proprietary” technologies this company says they provide?
a.
b.
Where is the fluorescent tag bound to the nucleotide? Why does this provide an improvement
over Sanger sequencing?
The sellers of this technology claim three improvements: Faster sequencing, longer read length,
and high fidelity.
a. What does “longer read length” likely mean?
B. What does “high fidelity” likely mean?
What is a ZMW? How would you describe it to a friend?
How many bases per second can the DNA polymerase add to the template?
Lecture 21 Page 3
How many ZMWs can run simultaneously? Circle on: (Dozens, Hundreds, Thousands, Millions)
The makers claim that eventually a researcher can sequence an entire genome in:
Time: ______________________ for Cost: _____________________
(Instructor disclaimer – This is one of many types of DNA sequencers. I’ve included it because
the biology is most relevant to our class – not because it is the best or cheapest. No need to rush
out and use this particular type).
21.5 Transcriptomics
Types of RNA:
Removal of rRNA:
Lecture 21 Page 4
Isolation of mRNA:
Sequencing of nucleotides:
21.6 Proteomics
Not all of the genome
Presence of mRNA…
Lecture 21 Page 5
Protein Mass Spectroscopy:
The spectrum of the molecule masses in graph is matched by computer to a database, to
produce a reading of most likely amino acids.
The sequence is matched to likely known proteins in a database for a final list of proteins found
in the cell.
Lecture 21 Page 6