Tentative
Lecture Schedule (Class room: Hurty Hall C in Fesler Hall)
|
6/4 Mon |
8:30 10:30 11:30 13:30 |
Introduction, DNA and RNA structures (Lee) DNA replication and growing E. coli for plasmid isolation (Lee) Transcription (Lee) Translation (Lee) |
|
6/5 Tue |
8:30 9:30 10:30 |
Genomic DNA isolation (Lee) Plasmid DNA isolation (Konger) Enzymes used in molecular biology (2 hours, Lee) |
|
6/6 Wed |
8:30 9:30 11:30 |
Enzymes used in molecular biology (Lee) Promoters and operons (2 hours, Lee) Total and Poly-A RNA isolation (Murrell) |
|
6/7 Thr |
8:30 9:30 10:30 11:30 |
Gel electrophoresis (Lee) Cloning vectors (Lee) Recover DNA from gels, Spin dialysis, DNA
concentration determination (Lee) Ligation, transformation, and screening of
recombinant plasmids (Lee) |
|
6/8 Fri |
8:30 9:30 10:30 |
Cloning strategies (Lee) E. coli genotypes
(Lee) DNA labeling and RNA probes (Lee) Southern and Northern (Lee) |
|
|
|
|
|
6/11 Mon |
8:30 9:30 10:30 11:30 |
Probe design
and microarrays (2 hrs, Lee) Library (1 hr) Restriction mapping (0.5 hr,
Lee) EMBL 3, EMBL4, Cosmid
(0.5 hr, Lee) |
|
6/12 Tue |
8:30 10:00 |
YAC, BAC, PAC (0.5 hours, Lee) PCR theory and
applications (3.5 hours) |
|
6/13 Wed |
8:30 9:30 11:30 |
PCR theory and applications (2 hr,
Lee) DNA sequencing principle (1 hr,
Lee) Biology of M13 and phagemid
(Lee) |
|
6/14 Thr |
8:30 9:30 10:30 |
Revolution of Sanger sequencing (Lee) In vivo excision of pBluescript
from lambda ZAP (0.5 Lee) Second generation sequencing techniques (2.5 hours,
Lee) |
|
6/15 Fri |
8:30 9:30 10:30 |
Expression of foreign genes in E. coli (Lee) Expression of foreign genes in eukaryotic
expression systems (Lee) Phage display, Tet-on/Tet-off and Gateway expression systems (2 hrs, Lee) |
|
|
|
|
|
6/18 Mon |
8:30 9:30 10:30 11:30 |
Mutagenesis and homologous recombination (Lee) Small and long non-coding RNA and RNA interference
(Lee) Proteomics (Wang) Fluorescent in situ hybridization |
|
6/19 Tue |
8:30 10:30 |
Viral vectors (2 hours, Lee) Bioinformatics and application of next generation
DNA sequencing (2 hours, Liu) |
|
6/20 Wed |
8:30 9:30 10:30 11:30 |
Molecular genetics (Dlouhy) Clinical applications of gene transfer technologies
(Cornetta) Transgenic animals and gene knockouts (Field) Single nucleotide polymorphism (Lee) |
|
6/21 Thr |
8:30 9:30 |
Single nucleotide polymorphism (Lee) Epigenetics (3 hours, Lee) |
Tentative Lab
Schedule
|
|
June 4 Mon |
June 5 Tue |
June 6 Wed |
June 7 Thr |
June 8 Fri |
|
Plasmid DNA |
1 ml-10 ml 10 ml-1 liter amplify (p8) Steps 2-4 |
Lyse cells Isolate DNA (p8-9) Steps 5-18 |
CsCl spin (p9-10) Steps 19-28 |
Purify band Dialyze (p10-11) Steps 29-37 |
Determine conc. Run gel (p11) Steps 38-40 |
|
Cloning |
|
|
Cut pMDM2 & vectors (p16) Electrophoresis (p17) |
Extract insert (p17-19) Ligate (p22-23) |
Transform (p23-25) |
|
Genomic DNA Southern |
Lyse cells Prot. K (p36) Steps 1-4 |
Extract DNA (p36-37) Steps 5-12 |
Purify & cut DNA (p37) Steps 13-16 |
Run gel (p37) Steps 17-18 |
Process gel & transfer (p38) |
|
Northern |
|
|
|
Isolate RNA (p45) Steps 1-12 |
Purify RNA (p45) Steps 13-17 |
|
|
June 11 Mon |
June 12 Tue |
June 13 Wed |
June 14 Thr |
June 15 Fri |
|
Cloning |
pTZ-pick, grow (p25) pACYC-rep. plate (p26) pBR-rep. plate (p28) |
Screen (pTZ) Grow (pACYC) Colony hyb. (pBR) |
Map pTZ recom. (p26) Screen pACYC recomb (p26) Hyb. (pBR; 29) |
Map pACYC recom. (p27) Wash Filters (pBR; p29) |
Develop film |
|
Southern |
Bake membrane (p38) |
Prehyb. (p40) |
Hyb. (p40) |
Wash, ARG (p40) |
Develop film |
|
Northern |
Run gel (p47) Transfer (p48) |
Bake Prehyb. |
Make probe hyb. |
Wash; ARG |
Develop film |
|
PCR/TOPO Cloning |
|
|
Isolate
template
(p52-54) |
PCR (p54-55) |
Ligate Transform
(p59-60) |
|
PCR/RFLP |
|
|
|
|
Spin dialyze DdeI digestion Run gel (p58) |
|
PCR/Dot-blot |
|
|
|
|
Denature Dot-blot (p55-57) Bake membrane |
|
|
June 18 Mon |
June 19 Tue |
June 20 Wed |
June 21 Thr |
|
PCR/Dot-blot |
Prehyb. (p57) |
Hyb. (p57) |
Wash & ARG (p57) |
Read results |
|
Real-time PCR |
|
Induce MIP-2, total RNA isolation,
reverse transcription (p63-64) |
PCR setup, Temp. cycling
(p64-65) Interpretation (p66) |
|
|
Luciferase Reporter
Assay |
Transfection (p68) |
Induction
(p68) |
Lyse cells;
determine luciferase activity (68-69) |
|