Making deletion crosses

DrosDel

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RS Elements
Molecular Biology
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Confirming Df's
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ED Stock Kit Available from Bloomington and Szeged Stock Centers:

You can download a PDF of the fly cross protocol for easier printing here.

StockDescriptionDrosDel CodeBloomington code
Isogenic stock w1118 iso; 2iso; 3iso DSK001 5905
Chr 2 balancer w1118 iso/Dp(1;Y)y+iso; nocSco/SM6a; 3iso DSK003 5907
Chr 2 balancer w1118 iso/Dp(1;Y)y+iso; wgGla/CyO; 3iso DSK019
Chr 3 balancer w1118 iso/Dp(1;Y)y+iso; 2iso/2iso; TM2/TM6C, Sb DSK004 5906
Chr 4 Balancer w1118 iso/Dp(1;Y)y+iso; 2iso/2iso;3iso; ciD (or eyD)/+ available from Cambridge
X balancer FM7h/P{RS3}l(1)CB-6411-3[1]; 2iso; 3iso DSK014 6878
X balancer Binsinscy/P{RS3}l(1)CB-6411-3[1]; 2iso; 3iso DSK018
FLP y w P{70FLP, ry+}3Fiso; 2iso; 3iso DSK009 6420
FLP for chr 2 deletions y w P{70FLP, ry+}3Fiso/y+Yiso; nocSco/SM6a; 3iso DSK010 6416
FLP for chr 3 deletions y w P{70FLP, ry+}3Fiso/y+Yiso; 2iso; TM2/TM6C, Sb DSK011 6419
X flippase source for Chr 4 deletions y w P{70FLP, ry+}3Fiso; 2iso; 3iso ; ciD (or eyD)/+ available from Cambridge
FLP for X deletions w1118iso; nocSco/SM6B, P{70FLP ry+}; 3iso DSK008 6876


Flip Out       RS w+ --> w-

Chromosome 2

™ y w 70FLPiso; Sco/SM6a; 3iso X ¡ w iso; P{FRT, w+}RS; 3iso -> heat shock
¡ y w 70FLPiso; P{FRT, w+}RS/SM6a; 3iso X ™ y w 70FLPiso; Sco/SM6a; 3iso->
single w- ¡ y w 70FLPiso; P{FRT, w-}RSr/SM6a; 3iso X ™ y w 70FLPiso; Sco/SM6a; 3iso -> stock

Chromosome 3

Same as 2, but using y w 70FLPiso; 2iso; TM2/TM6C; to balance.

Chromosome X

¡ w1118 iso; Sco/SM6b, 70FLP; 3iso X ™ w1118 P{FRT, w+}RS; 2iso; 3iso -> heat shock
¡ w1118 P{FRT, w+}RS; SM6b, 70FLP/2iso; 3iso X ™ FM7iso; 2iso; 3iso ->
single non-virgin w- FM7/w1118 P{FRT, w-}RSr; 2iso; 3iso X ¡ FM7iso; 2iso; 3iso ->
¡ w1118 P{FRT, w-}RSr; 2iso; 3iso X ™ FM7/ w1118 P{FRT, w-}RSr; 2iso; 3iso -> stock

Chromosome 4

™ y w 70FLPiso; 2iso; 3iso; ciD/eyD X ¡ w1118 iso; 2iso; 3iso; P{FRT, w-}RS -> heat shock
¡ y w 70FLPiso; 2iso; 3iso; P{FRT, w+}RS/ciD X ™ y w 70FLPiso; 2iso; 3iso; ciD/eyD ->
single w- ¡ y w 70FLPiso; 2iso; 3iso; P{FRT, w-}RSr/ciD X ™ y w 70FLPiso; 2iso; 3iso; ciD/eyD -> stock


Flip In      RS3r, w-/RS5r, w- --> Df w+

Chromosome 2

y w 70FLPiso; P{FRT, w-}RS3r; 3iso X y w 70FLPiso; P{FRT, w-}RS5r; 3iso -> heat shock
¡ y w 70FLPiso; P{FRT, w-}RS3r/ P{FRT, w-}RS5r; 3iso X ™ w1118 iso; Sco/SM6a; 3iso ->
single w+ ¡ w1118 iso; P{FRT, w+}RS3r--RS5r/SM6a; 3iso X ™ w1118 iso; Sco/SM6a; 3iso -> stock

Chromosome 3

Same as 2, but using w1118iso; 2iso;TM2/TM6C to balance.

Chromosome X

¡ w1118 iso; Sco/SM6b, 70FLP; 3iso X ™ w1118 P{FRT, w-}RS3r; 2iso; 3iso ->
¡ w1118 P{FRT, w-}RS3r; SM6b, 70FLP/2iso; 3iso X ™ w1118 P{FRT, w-}RS5r; 2iso; 3iso -> heat shock
™ w1118 P{FRT, w-}RS3r/w1118 P{FRT, w-}RS5r; SM6b, 70FLP/2iso; 3iso X ¡ FM7i iso; 2iso; 3iso ->
single w+ ™ w1118 P{FRT, w+}RS3r--RS5r/FM7i; 2iso; 3iso X ¡ FM7i iso; 2iso; 3iso -> stock

Chromosome 4

y w 70FLPiso; 2iso; 3iso; P{FRT, w-}RS3r/ciD X y w 70FLPiso; 2iso; 3iso; P{FRT, w-}RS5r/ciD -> heat shock
y w 70FLPiso; P{FRT, w-}RS3r/P{FRT, w-}RS5r; 3iso X w1118; 2iso; 3iso; ciD/eyD ->
single w+ ¡ w1118 iso; 2iso; 3iso; P{FRT, w+}RS3r--RS5r X ™ w1118; 2iso; 3iso; ciD/eyD -> stock

Heat shock protocol

Flip out: Heat shock for 1 hour at 37oC 48-72 hours after egg-laying (AEL).

Flip in: Heat shock for 1 hour at 37oC 48-72 hours AEL.
It may be necessary to heat shock again 48 hours later when making deletions on the X or larger autosomal deletions (>600kb).

References

Golic, K.G., Golic, M.M. Engineering the Drosophila genome: chromosome rearrangements by design. Genetics 1996 144(4):1693--1711