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Protease Inhibitors

Protease Inhibitors from Alberts' Lab Protocols (Kathy Miller/Chris Field)Protease inhibitors are rapidly inactivated in aqueous solutions and should be added to solutions immediately before use. Cock ...

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Basic Methods of Culturing Drosophila

Stockkeeping1. MechanicsMost stocks can be successfully cultured by periodic mass transfer of adults to fresh food. Bottles or vials are tapped on the pounding pad to shake flies away from the plug t ...

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How do you culture your Drosophila cells

I. MEDIUMA. S2 S2C Sundefined S2R+ S3 l(2)mbn Kc(167) & DL2 cells: Schneider's/ 10% FBS/ PS450 ml Schneider's medium (Gibco #11720-034)(pour off 50 ml into Falcon to save as serum-free~undefined*50_ml_Fetal_Bovine__...

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Current Bloomington Recipe for Drosophila Medium

This is our modification of a recipe used by John Loera at the University of Texas at Austin. The Texas recipe is apparently based on the malt medium described by Lakovaara (1969) with the addition ...

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Cuticle preparations

Cuticle preparationsThis procedure was adapted from one described over the phone (thanks P.G.). We're not sure if it appears elsewhere in published form.-Collect embryos on apple juice-agar plates for ...

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GENERAL MAINTENANCE INSTRUCTIONS FOR IMAGINAL DISC CELL LINES

GENERAL MAINTENANCE INSTRUCTIONS FOR IMAGINAL DISC CELL LINES This is our lab protocol for growing imaginal disc cells (clone-8) The Milner Lab website has additional protocols for clone-8 cells. Cl ...

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Hard Agar Medium for Egg Collection Plates

This is a firm medium that keeps well and makes a good base for egg collecting. Wrap filled petri dishes and store at 4-- 14oC (they should keep for at least 6 weeks). Substitute molasses for the c ...

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Sporulation in Liquid Media

Sporulation in Liquid Media1) Grow cells in 25mls of PSP2 (in 250ml flask) supplemented with 25% of the recommended concentration of amino acids until they are about 1x107 cells/ml (~1 day).2) Wash wi ...

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YPD FILLED MICROTITER PLATES

YPD FILLED MICROTITER PLATES Uses: 1. Grow individual strains to use for PCR2. Shipping strains 1. Make YPD regular agar (1.5-2%) for sending strains.2. Make 200ml batches = enough for 6 microtiter pl ...

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HC Dropout Plate

HC Dropout Plate Supplementation TableAmino AcidStockAmount To Spread(per 25 ml plate)URA1 mg/ml875 µlADE1 mg/ml500 µlLEU20 mg/ml100 µlLYS10 mg/ml300 µlMET2 mg/ml250 µlHIS10 mg/ml50 µlTRP10 mg/ml200 µ ...

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Methods for the preparation of Fly Extract (FE1 and FE2).

Fly extract 2 for routine culture (FE2) 1. First collect your flies; tip them from culture bottles into an empty glass bottle and plug it. Put it in the freezer for 45 mins or so until the flies are ...

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Production of Conditioned Medium containing Wingless protein from S2 cells

Production of Conditioned Medium containing Wingless protein from S2 cells- Grow heatshock wingless S2 cells at 25 degrees centigrade in a tissue culture flask to approximately 2.5 x 106 cells/ml.- He ...

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Routine methods for growing the cell lines

Medium Shields and Sang Medium 3 (SS3) modified for low serum. Supplemented before use with 2% heat inactivated FCS 2.5% FE2 12.5IU/100ml insulin and filter-sterilised through 0.22µm filter. Supplemen ...

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Methods for storing the cells in liquid nitrogen

Harvest cells and count as for passaging. 1x 107 cells in 1 ml are required for each 1 ml cryovial in 10% DMSO.For example:If 1ml cell suspension obtained from harvest contains 50 x 106 cells/ml.:For ...

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Labelling Drosophila pupae with BrdU

Labelling Drosophila pupae with BrdU ITO Kei 1992 OctoberCOLLECTING STAGED PUPAE 1. Prepare a couple of bottles containing late 3rd instar larvae which wander on the botlle wall (110-120 hours after e ...

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Single and Double FISH protocols for Drosophila

Single and Double FISH protocols for DrosophilaThis procedure is taken from a recently published book chapter. The reference is: Hughes S. and Krause H.M. (1998) Single and double FISH protocols for D ...

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Sporulation Method for BY4743.v1

Efficient sporulation of BY4743 requires growth as a patch on freshly prepared GNA presporulation plates at 30°C overnight prior to transfer to liquid sporulation medium. (Note: if GNA is inoculated ...

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Freezedown Guidelines

Freezedown Guidelines What to SaveFor each 96-pack each lab will have yeast representatives from all four strains: BY4741 (or BY4730) BY4742 (or BY4739) BY4743 and the homozygous diploid. It is recogn ...

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FEDEX Protocol

FEDEX Protocol(Or: How to increase the likelihood that your FEDEX packages reach their intended destination in a timely fashion)While this list seems pedantic so far it only incorporates actual bad ex ...

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Alternate Sporulation Method for BY4743

Alternate Sporulation Method for BY4743 Protocol: 1. Patch cells that have been freshly grown on YPD to freshly prepared GNA pre-sporulation plates for 1 day at 30°C.Repatch to another freshly prepar ...

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