Drosofilia Raccolta e dechorionation delle uova: un metodo per rimuovere lo strato di uova più esterno

Published: April 30, 2023

Abstract

Fonte: Koyle, M. L., et al. J. Vis. Exp. (2016).

L’embrione Drosophila è circondato da membrane esterne protettive. Per ottenere l’accesso all’embrione, il corione- il più esterno delle membrane – viene spesso rimosso. Questo video descrive un metodo comunemente usato di dechorionation usando ipoclorito di sodio. Il protocollo di esempio dimostra la procedura utilizzando tecniche sterili necessarie per l’allevamento animale axenico o gnotobiotico.

Protocol

Questo protocollo è un estratto da Koyle et al., Allevamento della Mosca della Frutta Drosophila melanogaster Sotto condizioni axeniche e gnotobiotiche, J. Vis. Exp. (2016). 1. Preparare gabbie per la deposizione delle uova Preparare piatti di agar succo d’uva microwaving 100 ml di acqua, 10 g di lievito di birra, 10 g di glucosio e 1 g di agar. Portare a ebollizione 3 volte nel passaggio 1.1 e aggiungere 10 g di concentrato di succo d’uva congelato per au…

Materials

Brewer's Yeast MP Biomedicals, LLC. 903312 http://www.mpbio.com/product.php?pid=02903312
Glucose Sigma Aldrich 158968-3KG http://www.sigmaaldrich.com/catalog/product/aldrich/158968?lang=en&region=US
Agar Fisher–Lab Scientific fly802010 https://www.fishersci.com/shop/products/drosophila-agar-8-100mesh-10kg/nc9349177
Welch's 100% Grape Juice Concentrate Walmart or other grocery store 9116196 http://www.walmart.com/ip/Welch-s-Frozen-100-Grape-Juice-Concentrate-11.5-oz/10804406
Cage: 32 oz. Translucent Round Deli Container Webstaurant Store 999L5032Y http://www.webstaurantstore.com/newspring-delitainer-sd5032y-32-oz-translucent-round-deli-container-24-pack/999L5032Y.html
Translucent Round Deli Container Lid Webstaurant Store 999YNL500 http://www.webstaurantstore.com/newspring-delitainer-ynl500-translucent-round-deli-container-lid-60-pack/999YNL500.html
Stock Bottles Genesee Scientific 32-130 https://geneseesci.com/shop-online/product-details/?product=32-130
Droso-Plugs Genesee Scientific 49-101 https://geneseesci.com/shop-online/product-details/?product=49-101
Nylon Mesh Genesee Scientific 57-102 https://geneseesci.com/shop-online/product-details/715/?product=57-102
Plastic Bushing Home Depot 100404002 http://www.homedepot.com/p/Carlon-1-in-Non-Metallic-Terminal-Adapter-E943FR-CTN/100404002
Plastic Bushing Cap Home Depot 100153897 http://www.homedepot.com/p/1-1-4- in-Rigid-Plastic-Insulated-Bushing-2-Pack-27529/100153897
Specimen Cup MedSupply Partners K01-207067 http://www.medsupplypartners.com/covidien-specimen-containers.html
Repeater M4 Eppendorf 4982000322 https://online-shop.eppendorf.us/US-en/Manual-Liquid-Handling-44563/Dispensers–Burettes-44566/Repeater-M4-PF-44619.html
50 ml Centrifuge Tubes TrueLine Centrifuge Tubes TR2003 https://www.lpsinc.com/Catalog4.asp?catalog_nu=TR2003
Food Boxes USA Scientific 2316-5001 http://www.usascientific.com/search.aspx?find=2316-5001
Lysing Matrix D Bulk MP Biomedicals, LLC. 116540434 http://www.mpbio.com/search.php?q=6540-434&s=Search
Filter Pipette Tips, 300 μl USA Scientific 1120-9810 http://www.usascientific.com/search.aspx?find=1120-9810
Petri Dishes Laboratory Product Sales M089303 https://www.lpsinc.com/Catalog4.asp?catalog_nu=M089303
Ethanol Decon Laboratories, INC. 2701 http://www.deconlabs.com/products.php?ID=88
Paintbrush Walmart 5133 http://www.walmart.com/ip/Chenille-Kraft-5133-Acrylic-Handled-Brush-Set-Assorted-Sizes-colors-8-Brushes-set/41446005
Forceps Fisher 08-882 https://www.fishersci.com/shop/products/fisherbrand-medium-pointed-forceps-3/p-128693
Household Bleach (6-8% Hypochlorite) Walmart 550646751 http://www.walmart.com/ip/Clorox-Concentrated-Regular-Bleach-121-fl-oz/21618295
Universal Peptone Genesee Scientific 20-260 https://geneseesci.com/shop-online/product-details/?product=20-260
Yeast Extract  Fisher Scientific BP1422-500 https://www.fishersci.com/shop/products/fisher-bioreagents-microbiology-media-additives-yeast-extract-3/bp1422500?matchedCatNo=BP1422500
Dipotassium Phosphate Sigma Aldrich P3786-1KG http://www.sigmaaldrich.com/catalog/
Ammonium Citrate Sigma Aldrich 25102-500g http://www.sigmaaldrich.com/catalog/
Sodium Acetate VWR 97061-994 https://us.vwr.com/store/catalog/product.jsp?catalog_number=97061-994
Magnesium Sulfate Fisher Scientific M63-500 https://www.fishersci.com/shop/products/magnesium-sulfate-heptahydrate-crystalline-certified-acs-fisher-chemical-3/m63500?matchedCatNo=M63500
Manganese Sulfate Sigma Aldrich 10034-96-5 http://www.sigmaaldrich.com/catalog/
MRS Powder Sigma Aldrich 69966-500G http://www.sigmaaldrich.com/catalog/product/sial/69966?lang=en&region=US
96 Well Plate Reader BioTek (Epoch) NA http://www.biotek.com/products/microplate_detection/
1.7 ml Centrifuge Tubes USA Scientific 1615-5500 http://www.usascientific.com/search.aspx?find=1615-5500
Filter Pipette Tips, 1,000 μl USA Scientific 1122-1830 http://www.usascientific.com/search.aspx?find=1122-1830
96 Well Plates Greiner Bio-One 655101 https://shop.gbo.com/en/usa/articles/
Ceramic Beads MP Biomedicals, LLC. 6540-434 http://www.mpbio.com/product.php?pid=116540434
Tissue Homogenizer MP Biomedicals, LLC. 116004500 http://www.mpbio.com/product.php?pid=116004500
Class 1 BioSafety Cabinet Thermo Scientific Model 1395 http://www.thermoscientific.com/en/product/1300-series-class-ii-type-a2-biological-safety-cabinet-packages.html

Play Video

Citazione di questo articolo
Drosophila Egg Collection and Dechorionation: A Method to Remove the Outermost Egg Layer. J. Vis. Exp. (Pending Publication), e20119, doi: (2023).

View Video