this week continue arabidopsis development screen, –discussion, –assignment (20 pts., submit one...
Post on 21-Dec-2015
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This Week
• Continue Arabidopsis Development Screen,
– discussion,
– assignment (20 pts., submit one per group),• describe your putants, describe and illustrate the growth and
development, through week 10 (due December 7th),– submit seeds in a sealed envelope (i.e., pick a fertile putant),
• Continue HFR experiment.
ScreensMulti-Quarter Project
Conditional Screen
• Rescue putants,• Complementation,• If known, submit to
Arabidopsis Biological Resource Center.
• If novel, map, then submit.
Developmental Screen
• Collect seeds,• Retest progeny,
– statistical relevance,
• Cross with wt,– F1, F2 segregation?
• Map,• Clone?
Classical Mapping
Cross an organism with a trait of interest to homozygous mutants of known mapped genes.
Then, determine if segregation is random in the F2 generation,
• if not, then your gene is linked (close) to the known mapped gene.
target
What recombination frequency do you expect between the target and HY2?
What recombination frequency do you expect between the target and TT2?
ScreensMulti-Quarter Project
Conditional Screen
• Rescue putants,
• Complementation,
• If known, submit to Arabidopsis Biological Resource Center.
• If novel, map, then submit.
Developmental Screen
• Collect seeds,• Retest progeny,
– statistical relevance,
• Cross with wt,– F1, F2 segregation?
• Map,• Clone?
Assignment(due December 7th)
– assignment (20 pts., submit one per group),
• describe your putants, describe and illustrate the growth and development, through week 10,
– dates, description, drawings, etc.,– special conditions, etc.
• submit seeds in a sealed envelope (i.e., pick fertile putants),
– staple envelopes to your report
lac (7)
car::tn10 (0.6)tetR
car (0.6)tetS
rpoB- (90)rifR
rpoB+ (90)rifS
metB- (89)
metB+ (89)
cysG- (72) cysG+ (72)
gyrA- (48)
nalS
gyrA+ (48)
nalR
1st Selection: tetR, nalR
Plate A proline, arginine, uracil
Plate B proline, arginine, uracil,methionine
Plate C proline, arginine, uracil,cysteine
Plate D proline, arginine, uracil,methionine, cysteine
Supplements Marker Selected
met, cys
cys
met
no selection
Selection Scheme
lac (7)
car::tn10 (0.6)tetR
car (0.6)tetS
rpoB- (90)rifR
rpoB+ (90)rifS
metB- (89)
metB+ (89)
cysG- (72) cysG+ (72)
gyrA- (48)
nalS
gyrA+ (48)
nalR
Met +, Cys + Met +, Cys - Met -, Cys +Met -, Cys -
Met+ Cys+ Plates: A __ B __ C __ D __
lac (7)
car::tn10 (0.6)tetR
car (0.6)tetS
rpoB- (90)rifR
rpoB+ (90)rifS
metB- (89)
metB+ (89)
cysG- (72) cysG+ (72)
gyrA- (48)
nalS
gyrA+ (48)
nalR
Met+ Cys- Plates: A __ B __ C __ D __Met- Cys+ Plates: A __ B __ C __ D __Met- Cys - Plates: A __ B __ C __ D __
lac (7)
car::tn10 (0.6)tetR
car (0.6)tetS
rpoB- (90)rifR
rpoB+ (90)rifS
metB- (89)
metB+ (89)
cysG- (72) cysG+ (72)
gyrA- (48)
nalS
gyrA+ (48)
nalR
Met_, Cys+ Plates: A __ B __ C __ D __
lac (7)
car::tn10 (0.6)tetR
car (0.6)tetS
rpoB- (90)rifR
rpoB+ (90)rifS
metB- (89)
metB+ (89)
cysG- (72) cysG+ (72)
gyrA- (48)
nalS
gyrA+ (48)
nalR
Met_, Cys- Plates: A __ B __ C __ D __