Smallmoleculeprep-1.growupwormssuperdense 2.spinwormsout 3.filtersupethroughwhatmanpaperandthenthroughsterilefilter 4.boilsupedownina2literflasktoabout50ml 5.spinputinsolublejunk,filterandboilto10ml 6.pinputjunkandboiltodryness-yellowishcrust 7.extractitwith10mlethanolovernightafterbreakingitupwithaspatulaintolittleyellowcrusts 8.repeatwithseveralO/Nextractions 9.speedvactheetohtodryness 10.resUSPendin10mlwaterandfreeze Cleanerprep:butloweryield1.growupamessawormsfrom1literculture 2.spinoutwormsandkeepthem 3.floatthemthroughsucrosetogetridofjunk 4.putwormsin250mlflaskwith50mls-basalandleaveat20degreesinshakero/n 5.spinoutworms 6.filtersupeandfreezeDauerPheromonePrep
Authors:FromCoriBargmannandBrendaReinhart1992AvariationofRiddle"sprotocol:1literofcultureofwormsins-basalande.coligrowfor7-10days,lettingitclearonce,feeditmoree.coliandletitclearagain
Note:whenwashworms,beverycarefultoavoidcontaminationandwashE.coliins-basaloncebeforeyouaddthem
