BISC219/F12: Lab 2: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
 
(8 intermediate revisions by 2 users not shown)
Line 2: Line 2:
<div style="padding: 10px; width: 720px; border: 5px solid #7E354D;">
<div style="padding: 10px; width: 720px; border: 5px solid #7E354D;">


== Lab 2:  Finish Series 1- Autosomal or Sex-linked? ==
== Lab 2:  Finish Project 1- Autosomal or Sex-linked Inheritance? ==
#Examine each plate of F2 progeny.  If you chose only L4 hermaphrodites, as instructed,you should only see hermaphrodite progeny.  If you have a lot of males on your plates, you probably chose young adult worms rather than L4 hermaphrodite's. That's a problem - see your instructor.<br>
#Examine each plate of F2 progeny.  If you chose only L4 hermaphrodites, as instructed,you should only see hermaphrodite progeny.  If you have a lot of males on your plates, you probably chose young adult worms rather than L4 hermaphrodite's. That's a problem - see your instructor.<br>
#For each cross, you should count and examine a random sample of 100 worms. The mutant worms may be smaller and not move as well as the wild type worms.  Look around your plate to get a quick assessment of the population.<br>
#For each cross, you should count and examine a random sample of 100 worms. The mutant worms may be smaller and not move as well as the wild type worms.  Look around your plate to get a quick assessment of the population.<br>
#Record in your lab notebook the number of WT, Dpy, Unc, or Dpy Unc mutants by examining the phenotype as you remove each animal from the plate. (Be sure to flame the pick after removing each worm!!!)  If the genes responsible for the mutations are unlinked, you should see WT's (+/+;+/+), Dpy’s (''dpy/dpy'';+/+), Unc’s(+/+;''unc/unc'') and Dpy Unc’s (''d/d;u/u'') in a ratio of 9:3:3:1.  If linked, you should see a greater proportion than expected of  Dpy Unc’s (''d u/d u'') double mutants vs Dpy or Unc single mutants among the mutant hermaphrodite progeny.<br>
#Record in your lab notebook the number of WT, Dpy, Unc, or Dpy Unc mutants by examining the phenotype as you remove each animal from the plate. (Be sure to flame the pick after removing each worm!!!)  If the genes responsible for the mutations are unlinked, you should see WT's (+/+;+/+), Dpy’s (''dpy/dpy'';+/+), Unc’s(+/+;''unc/unc'') and Dpy Unc’s (''d/d;u/u'') in a ratio of 9:3:3:1.  If linked, you should see a greater proportion than expected of  Dpy Unc’s (''d u/d u'') double mutants vs Dpy or Unc single mutants among the mutant hermaphrodite progeny.<br>
The main challenge is to correctly differentiate single mutants of each phenotype (Dpy or Unc) from double mutants(Dpy AND Unc). Students tend to undercount single mutants and overcount doubles when they are inexperienced. Check with your instructor if you are unsure about how to score these groups. WT worms may be undercounted because they are harder to "catch". Be careful not to skew your data in this way.  
The main challenge is to correctly differentiate single mutants of each phenotype (Dpy or Unc) from double mutants(Dpy AND Unc). Students tend to undercount single mutants and overcount doubles when they are inexperienced. Check with your instructor if you are unsure about how to score these groups. WT worms may be undercounted because they are harder to "catch". Be careful not to skew your data in this way. Pick what you consider single mutants to a plate and what you consider double mutants to another plate.  Ask your instructor to check them before you go to far in the scoring process.
'''Make sure you copy your data into the appropriate place on the course spreadsheet on the middle computer in the back of the lab!'''
'''Make sure you copy your data into the appropriate place on the course spreadsheet!'''
<br><br>
<br><br>
You should now be able to conclude which strain has mutations that are autosomal and linked, which strain has mutations that are both autosomal and unlinked, and which strain has an autosomal mutation and an x-linked mutation responsible for either the Dpy or Unc phenotype (which one?).<br>
You should now be able to conclude which strain has mutations that are autosomal and linked, which strain has mutations that are both autosomal and unlinked, and which strain has an autosomal mutation and an x-linked mutation responsible for either the Dpy or Unc phenotype (which one?).<br>
<br>
<br>
'''**See Assignments page for instructions on Series 1 Results section write up for this project. Due at the beginning of Lab 4.'''  [http://www.openwetware.org/wiki/BISC219/F12:_Assignment_Help-_Data_Analysis_1 Project 1 Write Up]
 
== Assignment ==
See [http://openwetware.org/wiki/BISC219/F12:_Assignment_Help-_Data_Analysis_1 | Assignment_Help-_Data_Analysis_1] for instructions on Series 1 Results section write up for this project. Due at the beginning of Lab 3.'''   
<div class=noprint>
<div class=noprint>


Line 28: Line 30:
[[BISC219/F12: Lab 7  | Lab 7: Complete Mapping: Score]]<br>
[[BISC219/F12: Lab 7  | Lab 7: Complete Mapping: Score]]<br>
'''Series3:'''<BR>
'''Series3:'''<BR>
[[BISC219/F12: RNA interference | Schedule of Reverse Genetics Project]]<BR>
[[BISC219/F12: RNAi General Information| Background Information on Project 3: Investigating Gene Regulation Using RNAi]] <br>
[[BISC219/F12: RNAi General Information | RNAi General Information]] <br>
[[BISC219/F12: Media Recipes | Media Recipes]]<br>
[[BISC219/F12: Media Recipes | Media Recipes]]<br>
[[BISC219/F12: RNAi Lab 5 | Lab 5: Picking your gene to RNAi]]<br>
[[BISC219/F12: RNAi Lab 7 | Lab 7: Identifying a bacterial colony containing our plasmid of interest  ]]<br>
[[BISC219/F12: RNAi Lab 6 | Lab 6: Cloning your gene of interest]]<br>
[[BISC219/F12: RNAi Lab 8 | Lab 8: Creating the feeding strain of bacteria for RNAi]]<br>
[[BISC219/F12: RNAi Lab 7  | Lab 7: Picking your transformant]]<br>
[[BISC219/F12: RNAi Lab 9 | Lab 9: Induction of feeding strain to produce dsRNA and feeding worms]]<br>
[[BISC219/F12: RNAi Lab 8 | Lab 8: Plasmid purification and transformation]]<br>
[[BISC219/F12: RNAi Lab 10 | Lab 10: Phenotypic analysis of treated vs untreated worms]]<br>
[[BISC219/F12: RNAi Lab | Lab 9: Induction of bacteria for RNAi]]<br>
[[BISC219/F12: RNAi Lab 11 | Lab 11: Writing Workshop]]<br>
[[BISC219/F12: RNAi Lab 10 | Lab 10: Scoring your worms]]<br>
[[BISC219/F12: RNAi Lab 12 | Lab 12: Writing Conferences]]<br>
[[BISC219/F12: RNAi Lab 11 | Lab 11: ]]<br><br>
</div>
</div>

Latest revision as of 08:53, 28 August 2012

Lab 2: Finish Project 1- Autosomal or Sex-linked Inheritance?

  1. Examine each plate of F2 progeny. If you chose only L4 hermaphrodites, as instructed,you should only see hermaphrodite progeny. If you have a lot of males on your plates, you probably chose young adult worms rather than L4 hermaphrodite's. That's a problem - see your instructor.
  2. For each cross, you should count and examine a random sample of 100 worms. The mutant worms may be smaller and not move as well as the wild type worms. Look around your plate to get a quick assessment of the population.
  3. Record in your lab notebook the number of WT, Dpy, Unc, or Dpy Unc mutants by examining the phenotype as you remove each animal from the plate. (Be sure to flame the pick after removing each worm!!!) If the genes responsible for the mutations are unlinked, you should see WT's (+/+;+/+), Dpy’s (dpy/dpy;+/+), Unc’s(+/+;unc/unc) and Dpy Unc’s (d/d;u/u) in a ratio of 9:3:3:1. If linked, you should see a greater proportion than expected of Dpy Unc’s (d u/d u) double mutants vs Dpy or Unc single mutants among the mutant hermaphrodite progeny.

The main challenge is to correctly differentiate single mutants of each phenotype (Dpy or Unc) from double mutants(Dpy AND Unc). Students tend to undercount single mutants and overcount doubles when they are inexperienced. Check with your instructor if you are unsure about how to score these groups. WT worms may be undercounted because they are harder to "catch". Be careful not to skew your data in this way. Pick what you consider single mutants to a plate and what you consider double mutants to another plate. Ask your instructor to check them before you go to far in the scoring process. Make sure you copy your data into the appropriate place on the course spreadsheet!

You should now be able to conclude which strain has mutations that are autosomal and linked, which strain has mutations that are both autosomal and unlinked, and which strain has an autosomal mutation and an x-linked mutation responsible for either the Dpy or Unc phenotype (which one?).

Assignment

See | Assignment_Help-_Data_Analysis_1 for instructions on Series 1 Results section write up for this project. Due at the beginning of Lab 3.