Plant Chromosome Preparation & FISH Workshop
May 14, 2012–May 18, 2012
Organized by:
- Nurul Islam-Faridi, PhD
Research Molecular Cytogeneticist (USFS)
Adjunct Professor (TAMU) - Forest Tree Molecular Cytogenetics Laboratory
- Southern Institute of Forest Genetics
Southern Research Station, U.S. Forest Service - Department of Ecosystem Science & Management Texas A&M University
| May 14 Monday | |
| 09:00 – 09:15 | Welcome address by Dr. Tom Byram, Director, Western Gulf Forest Tree Improvement Program, Texas Forest Service & Texas A&M |
| 09:15 – 10:00 | FISH presentation (Emphasis on proper plant chromosome spreading) |
| 10:10 – 11:00 | Root process for enzymatic digestion |
| 11:00 – 12:30 | Root collection – Greenhouse |
| 12:30 – 13:30 | Lunch (on your own) |
| 13:30 – 18:00 | RNase Pre-treatment for FISH Fixation root tips Root processing – Enzymatic digestion – Slide preparation |
| 18:00 – 19:30 | Dinner on your own |
| 19:30 – 22:00 | Video Demo: Metafer (MetaSystems) – Automated detection of metaphase spreads & karyotyping |
| May 15 Tuesday | |
| 08:30 – 17:30 | Practice on your own: Process roots Enzymatic digestion Slide preparation |
| 9:00 – 10:00 | FISH discussion – Methodology & Unforeseen problems |
| 12:30 – 13:30 | Lunch (on your own) |
| 13:30 – 17:30 |
FISH Day – 1 Hybridization mixture preparation Slide Denaturation (Chromosomal DNA) Hybridization (DNA to DNA) Overnight incubation Set-up for hybridization site detection (FISH Day -2) |
| May 16 Wednesday | |
| 08:00 – 11:30 | FISH Day – 2 Washing & Hybridization site(s) detection |
| 11:30 – 12:30 | Discussion: FISH with BAC, Transgene and genomic DNA probes; Pachytene FISH |
| 12:30 – 13:30 | Lunch (on your own) |
| 13:30 – 18:00 | Operation of epi-fluorescence microscope and use of MetaSystems FISH analysis software. We have two (State-of-the-Art)) epi-fluorescence microscopes (automated and manual) and will provide training for the both |
| May 17 Thursday | |
| 08:00 – 17:00 | Nick-Translation – Cloned DNA Check Labeled DNA on gel View FISH slides, capture and process FISH images |
| 12:30 – 13:30 | Lunch (on your own) |
| May 18 Friday | |
| 08:00 – 12:30 | FISH slide analysis – Microscope Metafer (MetaSystems Group Inc.) – automated detection of metaphase spreads |
| 12:30 – 13:30 | Lunch (on your own) |
| 13:30 – 16:00 | Capture & process FISH images |
| 16:00 – 17:30 | Question & Answer Session Adjourn |
PS:
Participants will be encouraged to submit a short report on the following headings within a week from May 18, 2012.
Plant somatic chromosome preparation (root tip collection, pretreatment, fixation, enzymatic digestion and chromosome spread) and FISH (probe DNA, probe labeling and detection, RNase-A treatment, hybridization mixture, denaturation, hybridization and detection, capturing and pressing FISH image. The report should include two/three FISH images).
For additional details please contact the workshop organizer.
E-mail: nfaridi@tamu.edu; telephone: 979-862-3908 (office), 979-845-2974 (lab).