Tentative Lecture Schedule
|
6/8 Mon |
8:30 10:30 11:30 13:30 |
Introduction,
DNA and RNA structure (Lee) DNA
replication and growing E. coli for
plasmid isolation (Lee) Transcription
(Lee) Genomic
DNA isolation and translation (Lee) |
|
6/9 Tue |
8:30 10:30 11:30 |
Translation
(2hrs, Lee) Plasmid
DNA isolation (Konger) Enzymes
used in molecular biology (Lee) |
|
6/10 Wed |
8:30 10:30 11:30 |
Enzymes
used in molecular biology (2 hrs, Lee) Promoters
and operons (Lee) Gel
electrophoresis (Lee) |
|
6/11 Thr |
8:30 9:30 10:30 11:30 |
Total
and Poly-A RNA isolation (Murrell) Recover
DNA from gels, Spin dialysis, DNA concentration determination (Lee) Cloning
vectors (Lee) Ligation,
transformation, and screening of recombinant plasmids (Lee) |
|
6/12 Fri |
8:30 9:30 10:30 |
Cloning
strategies (Lee) E. coli genotypes (Lee) DNA
labeling, DNA and RNA probes (2hrs, Lee) |
|
|
|
|
|
6/15 Mon |
8:30 9:30 10:30 11;30 |
Southern
and Northern (Lee) Hybridization
(Lee) Life
cycle of lambda phage (Lee) cDNA synthesis and cloning
(Lee) |
|
6/16 Tue |
8:30 9:30 10:30 |
Lambda
packaging and cDNA library screening (Lee) Restriction
mapping (Lee) DNA
sequencing (2hrs, Lee) |
|
6/17 Wed |
8:30 9:30 10:30 11;30 |
Biology
of M13 (Lee) Phagemid and in vivo excision of pBluescript
from lambda ZAP (Lee) EMBL
3, EMBL4, Cosmid, YAC, BAC, PAC (Lee) Proteomics
(Wang) |
|
6/18 Thr |
8:30 9:30 |
DNA
sequence analyses (Liu) PCR
theory and applications (3 hrs, Lee) |
|
6/19 Fri |
8:30 10:30 11:30 |
PCR
theory and applications (2 hrs, Lee) Functional
genomics (Xuei) Single
nucleotide polymorphism (Xuei) |
|
|
|
|
|
6/22 Mon |
8:30 10:30 11:30 |
Expression
of foreign genes in E. coli (2 hrs,
Lee) Expression
of foreign genes in eukaryotic expression systems (Vidal) Gateway
expression systems (Lee) |
|
6/23 Tue |
8:30 9:30 11:30 |
Fluorescent
in situ hybridization (Thurston) Mutagenesis
and homologous recombination (Lee) Viral
vectors for gene transfer (2 hrs, Lee) |
|
6/24 Wed |
8:30 9:30 10:30 11:30 |
Molecular
Genetics (Dlouhy) Molecular
diagnostics (Blue) Transgenic
animals and gene knockouts (Field) Clinical
applications of gene transfer technologies (Cornetta) |
|
6/25 Thr |
8:30 9:30 10:30 11:30 |
siRNA and ET/Red cloning system
(Lee) Transfection, reporter assays, gel shift, footprinting, 2-hybrid (Lee) Primer
extension, S1, RNaseA protection, SSCP, RAPD, AFLP
(Lee) mRNA differential display, DNA microarrays, TOGA,
SAGE, MPSS, SOLiD, RDA (Lee) |
Tentative Lab
Schedule
|
|
June 8 Mon |
June 9 Tue |
June 10 Wed |
June 11 Thr |
June 12 Fri |
|
Plasmid DNA |
1 ml-10 ml 10 ml-1 liter amplify (p10) Steps 2-4 |
Lyse cells Isolate DNA (p10-11) Steps 5-18 |
CsCl spin (p11-12) Steps 19-28 |
Purify band Dialyze (p12-13) Steps 29-37 |
Determine conc. Run gel Steps 38-40 |
|
Cloning |
|
|
Cut pMDM2 & vectors
(p18) Electrophoresis (p19-20) |
Extract insert Ligate (p19-28) |
Transform (p28-31) |
|
Genomic DNA Southern |
Lyse cells Prot. K (p44) Steps 1-4 |
Extract DNA (p44-45) Steps 5-12 |
Purify & cut DNA (p45) Steps 13-16 |
Run gel (p45) Steps 17-18 |
Process gel & transfer
(p46) |
|
Northern |
|
|
|
Isolate RNA (p53) Steps 1-12 |
Purify RNA (p53) Steps 13-17 |
|
|
June 15 Mon |
June 16 Tue |
June 17 Wed |
June 18 Thr |
June 19 Fri |
|
Cloning |
pACYC-rep. plate pTZ-pick,
grow pBR-rep.
Plate |
Grow Screen Colony hyb. |
Screen (p32-36) Map (p32-36) Hyb. (p32-36) |
Map Wash Filters |
Develop film |
|
Southern |
Bake membrane (p46) |
Prehyb. (p48) |
Hyb. (p48) |
Wash, ARG (p48) |
Develop film |
|
Northern |
Run gel (p55) Transfer (p56) |
Bake Prehyb. (p57) |
Make probe hyb.
(p57) |
Wash; ARG (p57) |
Develop film |
|
PCR/TOPO Cloning |
|
|
Isolate template
(p61-62) |
PCR (p62-63) |
Ligate Transform
(p66-67) |
|
PCR/RFLP |
|
|
|
|
Spin dialyze DdeI digestion Run gel (p65) |
|
PCR/Dot-blot |
|
|
|
|
Denature Dot-blot (p63-64) Bake membrane |
|
|
June 22 Mon |
June 23 Tue |
June 24 Wed |
June 25 Thr |
|
PCR/Dot-blot |
Prehyb.
(p64) |
Hyb. (p64) |
Wash (p64), ARG
(p64) |
Read results |
|
Real-time PCR |
|
Induce iNOS, total RNA isolation, reverse transcription (p71-72) |
Setup, Temp. cycling Interpretation
(p72-73) |
|