<em>Drosophila</em> Egg Collection and Dechorionation: A Method to Remove the Outermost Egg Layer

doi:10.3791/20119

JoVE Journal > Embryo

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Embryo

드로소필라 계란 컬렉션 및 불반향 : 가장 바깥쪽 계란 층을 제거하는 방법

Published: April 30, 2023

Abstract

출처: 코일, M. L., 외. 악센과 그노토바이오틱 조건 하에서 과일 플라이 드로소필라 멜라노가스터를 사육. J. 비스 익스펙. (2016).

드로소필라 배아는 보호적인 외부 막으로 둘러싸여 있습니다. 배아에 접근하기 위하여는, chorion- 막의 가장 바깥쪽 – 수시로 제거됩니다. 이 비디오는 나트륨 hypochlorite를 사용하여 일반적으로 사용되는 비초화 방법을 설명합니다. 이 예제 프로토콜은 축또는 노토바이오틱 동물 사육에 필요한 멸균 기술을 사용하는 동안 절차를 보여줍니다.

Protocol

이 프로토콜은 코일 외에서발췌, 악센과 노토바이오틱 조건에서 과일 플라이 드로소필라 멜라노가스터를 사육, J. Vis. Exp. (2016). 1. 계란 누워 케이지를 준비 포도 주스 한천 접시를 100ml 의 물, 맥주 효모 10g, 포도당 10g, 한천 1 g로 만듭니다. 1.1단계에서 3번 끓이고 냉동 포도 주스 농축액 10g을 첨가하여 한천 판에 달걀의 가시성을 높입니다. 한…

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

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