BISC219/F12:RNA interference

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(New page: {{Template:BISC219/F12}} <div style="padding: 10px; width: 720px; border: 5px solid #7E354D;"> These labs were developed with the help of the '''Silencing Genomes''' project of the Dolan ...)
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! Lab # !! Dates !! Activity !! Outside lab time
! Lab # !! Dates !! Activity !! Outside lab time
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! 5
 
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| 9/29 - 10/5
 
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| Pick a gene of interest<br>
 
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Set up a PCR reaction to clone the gene
 
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|Examine the results of the agarose gel electrophoresis
 
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!6
 
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|10/12 - 10/18
 
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|Restriction enzyme digest of PCR product<br>
 
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Cleanup and ligation into the pPD129.36 vector<br>
 
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Transformation of the isolated plasmid into the BL21 cloning ''E. coli''
 
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|'''The day after lab:'''<br>
 
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Check control and transformation plates for growth - save your transformation plate<br>
 
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!7
!7
|10/19 - 10/25
|10/19 - 10/25
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|Colony PCR to check for transformation
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|'''The night before next lab:'''<br>
|'''The night before next lab:'''<br>
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Set up an overnight culture of a single colony from your transformation
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!8
!8
|11/2 - 11/8
|11/2 - 11/8
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|Plasmid isolation from the BL21 cells <br>
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Quantification of DNA<br>
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Transformation of plasmid into the HT115(DE3) cells
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|'''The day after lab:'''<br>
|'''The day after lab:'''<br>
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Check control and transformation plates for growth - save your transformation plate<br>
 
'''The night before next lab:'''<br>
'''The night before next lab:'''<br>
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Set up an overnight culture of a single colony from your transformation
 
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!9
!9
|11/9 - 11/15
|11/9 - 11/15
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|Induction of the bacteria to produce RNA<br>
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Seed plates and dry for bacterial feeding RNAi<br>
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|'''4 days later:'''<br>
|'''4 days later:'''<br>
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Pick 2 L4 hermaphrodite worms of N2 and ''rrf-3'' genotype to 2 RNAi plates for each genotype and make 1 control "mock" plate for each genotype as well ('''6 plates total''')<br>
 
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Incubate at 23°C until next class
 
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!10
!10
|11/16 - 11/22
|11/16 - 11/22
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| Examine the phenotypes of the fed worms - compare to control N2 worms and worms containing a mutation in the gene you are examining<br>
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Collection of treated worms and RNA purification
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!11
!11
|11/28 - 12/2
|11/28 - 12/2
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|Sign up for a time to meet and discuss your final paper draft with your instructor - we will not meet in lab this week.
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!12
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|12/ - 12/
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Revision as of 15:19, 20 August 2012

Wellesley College BISC 219 Genetics

These labs were developed with the help of the Silencing Genomes project of the Dolan DNA Learning Center.

RNAi General Information
Media Recipes
Lab 5: Picking your gene to RNAi
Lab 6: Cloning your gene of interest
Lab 7: Picking your transformant
Lab 8: Plasmid purification and transformation
Lab 9: Induction of bacteria for RNAi
Lab 10: Scoring your worms
Lab 11: Individual Writing Conferences


Schedule of Experiments

Lab # Dates Activity Outside lab time
7 10/19 - 10/25 The night before next lab:
8 11/2 - 11/8 The day after lab:

The night before next lab:

9 11/9 - 11/15 4 days later:
10 11/16 - 11/22
11 11/28 - 12/2
12 12/ - 12/
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