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Welcome to MCDB 306
Genetics Laboratory
Nhu Do
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Lecture 01 January 11th
• Todays !utline – Course "olicies
– #ntroduction to"ro$ects andDrosophilamelanogaster
• %nnouncements – #& you are 'ait listed
&or a lab( "lease )o tothe lab you 'ant toattend and see i&there is s"ace
– We 'ill &ollo' 'ait listorder to *ll anyremainin) s"ots
Wee+ o& January 1,th and 'ee+
!& January -.th / ou 'ill need to come into lab mornin)and eenin) hours to collect 2ies
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#nstructors
• Lecturer
– Diane 4"illane 5ds"ill( 678-7-19• !:ce hours Thursday 1081-;00 "m( 13,< =4B
•
GSIs and their Ofce hours held in 3131 USB – Din) >e( Wednesday 10 am81;00 "m
– 4usanna Cam"bell( Wednesday .8,;00 "m
– 4urabhi ?a$aram( Thursday .8,;00 "m
–
Chris 4i&uentes( @riday 11 am8-;00 "m
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Gradin) olicy
Grade Scale
• 360800 "ts % ran)e
• 3-083.E "ts B ran)e
• -,0831E "ts C ran)e• -08-7E "ts D ran)e
• F -3E "ts <
• #& necessary( these cutoHs 'ill be cured Nocurin) is done until the com"letion o& thecourse
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Grades/00 oints Total
• Iuies – 100 "ts( -.K o& )rade
• Drosophila ro$ect
– 16 "ts( 1K o& )rade• C. elegans ro$ect
– .0 "ts( 1-.K o& )rade
• >ome'or+ – 60 "ts( 1.K o& )rade
• Laboratory 'or+ – -6 "ts( 6.K o& )rade
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Iuies
• There are t'o Iuies )ien durin) thelast hour o& your laboratory "eriod – @ebruary --8-3rd
– %"ril 8.th
• #& you reuire s"ecial accommodations(you must brin) the a""ro"riate
documentation to Dr 4"illane at least -'ee+s "rior to the *rst ui
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Course Canas 4ite
• Lab Manual –
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4ub$ect Matter
• Drosophila "ro$ect – #denti&y "otential re)ulators o& a cell8cell
communication "ath'ay 5Win)less or Wnt si)nalin)9• #denti&y the chromosomal location o& random trans"oson
insertions that aHect the Win)less "ath'ay
• =se C? &ollo'ed by DN% seuencin) to more "reciselyidenti&y the trans"oson insertion 'ith the )oal o& *ndin)"otential )enes inoled in the Win)less "ath'ay
• Caenorhabditis elegans 5C. elegans9"ro$ect – Genetic ma""in) o& a mutation usin) molecular
mar+ers
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>o' are 'e )oin) to identi&y re)ulators o& W) si)nalin)
1 4tart 'ith a "henoty"e that results &rom oereA"ressionWin)less
- % 4creen 'as "er&ormed to identi&ytrans"oson insertions that su""ress the"henoty"e
!ereA"ressiono& W) in the
eye causes asmall
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The search &or re)ulators o& W) si)nalin) usestrans"oson
5G4OP9 insertions that allo' nearby )enes to be
actiated in an inducible manner4tretch o& 2y)enomic DN%
#nsert a construct5"G4OP9
G4OP
We hae many diHerent 2y stoc+s 'ith random G4OP insertionsMore on this later
The insert causes transcri"tional actiation o&
)enes oneither side o& the insert( )enes that may be
inoled in the W) si)nalin) "ath'ay
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#ntroduction to Drosophila
melanogaster
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Why =se Drosophila
• 4mall sie
• 4hort li&e cycle 51081 days9
•
>i)h re"roductie rate 5a sin)le&emale can lay 008.00 e))s in 10days9
•
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Droso"hila Li&e Cycle
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Becomin) @amiliar 'ithDroso"hila
horaA has three se)ments( each 'ith a "air o& le)s The 'iattached to T-( the haltere to T3
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%bdominal 4e)ments in Drosophila
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Droso"hila Chromosomes
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Droso"hila chromosomes
4econd "air
Third "air
4eA chromosome
@ourth "air
Di"loid-n Q ,
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Drosophila Cultures s Drosophila
Crosses
• Cultures – Males and &emales are o& the same
)enoty"es
–Ree"s a stoc+ o& 2ies &or &uture uses
• Crosses – Males and &emales are o& diHerent
)enoty"es
– =sed &or eA"erimental "ur"oses
– ou 'ill uses crosses to ans'er uestionsabout your un+no'n insertions
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Males s @emales( cont
Males hae sexcombs on the *rst
set o& le)s#m"ortant &or matin)
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The Best Way to 4eA @lies
• The male genitalclasers are themost accurate 'ay todistin)uish a male&rom &emale
• This is 'hat youshould do in thelab
• Could con*rm 'ithseA combs( but ta+estoo much time
ou tube ideo; htt";'''youtubecom'atchQ&Sla&h@$ec
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4ymbolism =sed• Wild Ty"e
–
We use the symbol U to denote 'ildty"e
– Can 'rite U ty"e
• Mutant
– %ny heritable deiation &rom 'ild ty"e ismutant
– We use an abbreiation o& the name(
such as ' &or 'hite 5'hite eye colormutation9
– Lo'er case letter &or a recessiemutation
– = er case letter &or a dominant
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Nomenclature continued
• Commas are used to se"arate traitson the same chromosome
• 4emi8colons are used to se"aratetraits on diHerent chromosomes
'' UU LyTM6B( 4b
What does this mean
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'' UU LyTM6B( 4b
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'' UU LyTM6B( 4b
The recessie 'hitemutation is located onthe V chromosome
This is a &emale
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The semi8colon se"arates the chromosomes
'' UU LyTM6B( 4b
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'' UU LyTM6B( 4b
Ly is a dominant mutationlocated on one co"y o& the3rd chromosome
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'' UU LyTM6B( 4b
Ly is a dominant mutationlocated on one co"y o& the3rd chromosome
TM6B is a dominant mutationlocated on the other co"y o&
The 3rd chromosome
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'' UU LyTM6B( 4b
Ly is a dominant mutationlocated on one co"y o& the3rd chromosome
TM6B is a dominant mutationlocated on the other co"y o&
The 3rd chromosome
4b is a dominant mutationlocated on the other co"y o
The 3rd chromosome( thesame chromosome as TM6
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Lab This Wee+
• Learn ho' to "ro"erly use "i"ettors
• Collect 30 2ies to be used &or GenomicDN% isolation
• Learn ho' to handle Droso"hila – =se the C0- system to anesthetie 2ies
• "er# imortant$$$
– #denti&y 2y body "arts and sta)es o& the li&ecycle
– Become com%ortable searating malesand %emales$
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Will # be 'or+in) alone
• @or the Droso"hila ro$ect – ou 'ill 'or+ in "airs to collect data
5some control data 'ill be shared 'ith
the class9 – >o'eer( all 'ritten re"orts and
home'or+ must be done indiidually
•
@or the C ele)ans ro$ect – ou 'ill 'or+ indiidually to collect data
and com"lete re"orts and home'or+