BISC219/F12:RNA interference: Difference between revisions
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These labs were modified from the work of Janet Andersen, Alexander Krichevsky, Joerg R. Leheste, and Daniel J. Moloney at the Dept. of Biochemistry and Cellular BIology at Stony Brook University with | These labs were modified from the work of Janet Andersen, Alexander Krichevsky, Joerg R. Leheste, and Daniel J. Moloney at the Dept. of Biochemistry and Cellular BIology at Stony Brook University with the '''Silencing Genomes''' project of the Dolan DNA Learning Center in Cold Spring Harbour, New York. providing some of the worm strains.<br><br> | ||
[[BISC219/F12: RNAi General Information| RNAi | [[BISC219/F12: RNAi General Information| Background Information on Project 3: Investigating Gene Function & Regulation Using RNAi]] <br> | ||
[[BISC219/F12: Media Recipes | Media Recipes]]<br> | [[BISC219/F12: Media Recipes | Media Recipes]]<br> | ||
[[BISC219/F12: RNAi Lab 7 | Lab 7: ]]<br> | [[BISC219/F12: RNAi Lab 7 | Lab 7: Identifying a bacterial colony containing our plasmid of interest ]]<br> | ||
[[BISC219/F12: RNAi Lab 8 | Lab 8: ]]<br> | [[BISC219/F12: RNAi Lab 8 | Lab 8: Creating the feeding strain of bacteria for RNAi]]<br> | ||
[[BISC219/F12: RNAi Lab 9 | Lab 9: ]]<br> | [[BISC219/F12: RNAi Lab 9 | Lab 9: Induction of feeding strain to produce dsRNA and feeding worms]]<br> | ||
[[BISC219/F12: RNAi Lab 10 | Lab 10: ]]<br> | [[BISC219/F12: RNAi Lab 10 | Lab 10: Phenotypic analysis of treated vs untreated worms]]<br> | ||
[[BISC219/F12: RNAi Lab 11 | Lab 11: ]]<br><br> | [[BISC219/F12: RNAi Lab 11 | Lab 11: Data Analysis Workshop]]<br> | ||
[[BISC219/F12: RNAi Lab 12 | Lab 12: Optional Conferences]]<br> | |||
<br> | |||
| Line 20: | Line 22: | ||
!7 | !7 | ||
|10/23 - 10/29 | |10/23 - 10/29 | ||
| | | Picking a colony of bacteria containing the RNAi plasmid<br> | ||
|'''The night before next lab:'''<br> | PCR to confirm presence of gene of interest | ||
|'''The night before next lab:''' set up an overnight culture of your colony of interest<br> | |||
|- | |- | ||
!8 | !8 | ||
|10/30 - 11/5 | |10/30 - 11/5 | ||
| | | Miniprep of plasmid and transformation into feeding bacteria strain | ||
|'''The day after lab:'''<br> | |'''The day after lab:''' check your transformed bacteria for colony growth<br> | ||
'''The night before next lab:'''<br> | '''The night before next lab:''' set up an overnight culture of your colony of interest<br> | ||
|- | |- | ||
!9 | !9 | ||
|11/7 - 11/13 | |11/7 - 11/13 | ||
| Induction of dsRNA production, seeding of RNAi plates and addition of worms | |||
| | | | ||
|- | |- | ||
!10 | !10 | ||
|11/14 - 11/20 | |11/14 - 11/20 | ||
| | | Chemotaxis Assay | ||
| | | | ||
|- | |- | ||
!11 | !11 | ||
|11/26 - 11/ | |11/26 - 11/30 | ||
| | | Writing Workshop | ||
| | | | ||
|- | |- | ||
!12 | !12 | ||
|12/ - 12/7 | |12/3 - 12/7 | ||
| | | Writing Conferences | ||
| | | | ||
|- | |- | ||
Latest revision as of 18:04, 18 November 2012
These labs were modified from the work of Janet Andersen, Alexander Krichevsky, Joerg R. Leheste, and Daniel J. Moloney at the Dept. of Biochemistry and Cellular BIology at Stony Brook University with the Silencing Genomes project of the Dolan DNA Learning Center in Cold Spring Harbour, New York. providing some of the worm strains.
Background Information on Project 3: Investigating Gene Function & Regulation Using RNAi
Media Recipes
Lab 7: Identifying a bacterial colony containing our plasmid of interest
Lab 8: Creating the feeding strain of bacteria for RNAi
Lab 9: Induction of feeding strain to produce dsRNA and feeding worms
Lab 10: Phenotypic analysis of treated vs untreated worms
Lab 11: Data Analysis Workshop
Lab 12: Optional Conferences
Schedule of Experiments
| Lab # | Dates | Activity | Outside lab time |
|---|---|---|---|
| 7 | 10/23 - 10/29 | Picking a colony of bacteria containing the RNAi plasmid PCR to confirm presence of gene of interest |
The night before next lab: set up an overnight culture of your colony of interest |
| 8 | 10/30 - 11/5 | Miniprep of plasmid and transformation into feeding bacteria strain | The day after lab: check your transformed bacteria for colony growth The night before next lab: set up an overnight culture of your colony of interest |
| 9 | 11/7 - 11/13 | Induction of dsRNA production, seeding of RNAi plates and addition of worms | |
| 10 | 11/14 - 11/20 | Chemotaxis Assay | |
| 11 | 11/26 - 11/30 | Writing Workshop | |
| 12 | 12/3 - 12/7 | Writing Conferences |
