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    Tigriopus epigenetics miniproject

    In this exercise, you will test for changes in methylation of the coding sequence of heat shock protein 70 in response to heat shock in Tigriopus using restriction enzyme digests with MspI (methylation insensitive) and HpaII (methylation sensitive)

    DNA extraction and enzyme digest:

    1) remove as much water as possible from tube containing 1 copepod. Add 30 ul of chelex, vortex, spin down, incubate at 95˚C for 20 minutes.

    2) Vortex chelex solution again, spin down for 1 minute on high speed

    3) Put 5ul of DNA template into each of 3 pcr tubes – one for each enzyme and one for the ‘water only’ control.

    4) To each tube add: 0.5 ul enzyme (or milliQ water), 2ul of cutsmart buffer, 12 ul of water.

    5) Incubate at 37˚C for 2 hours, followed by 5 min at 95˚C.

    PCR:

    6) To each enzyme digest tube, add 0.2 ul Taq, 0.5ul Forward+Reverse HSP70 primer mix, 1.6 ul dNTPs

    7) PCR cycle: “EPI”

    Comments

    /groups/eccofhl/search/index.rss?tag=hotlist/groups/eccofhl/search/?tag=hotWhat’s HotHotListHot!?tag=hot4/groups/eccofhl/sidebar/HotListjohnwaresjohn wares2016-06-30 15:22:33+00:002016-06-30 15:22:33updated32johnwaresjohn wares2016-06-29 02:28:51+00:002016-06-29 02:28:51updated31johnwaresjohn wares2016-06-29 01:42:23+00:002016-06-29 01:42:23updated30johnwaresjohn wares2016-06-29 01:42:02+00:002016-06-29 01:42:02updated29johnwaresjohn wares2016-06-28 15:02:12+00:002016-06-28 15:02:12updated28johnwaresjohn wares2016-06-24 04:48:13+00:002016-06-24 04:48:13updated27Added tag - hotjohnwaresjohn wares2016-06-23 17:37:21+00:002016-06-23 17:37:21addTag26johnwaresjohn wares2016-06-22 23:47:31+00:002016-06-22 23:47:31updated25johnwaresjohn wares2016-06-21 18:50:55+00:002016-06-21 18:50:55updated24johnwaresjohn wares2016-06-21 02:38:03+00:002016-06-21 02:38:03updated23johnwaresjohn wares2016-06-21 02:36:50+00:002016-06-21 02:36:50updated22johnwaresjohn wares2016-06-21 02:31:29+00:002016-06-21 02:31:29updated21johnwaresjohn wares2016-06-21 02:18:48+00:002016-06-21 02:18:48updated20johnwaresjohn wares2016-06-19 16:28:21+00:002016-06-19 16:28:21updated19johnwaresjohn wares2016-06-19 16:27:22+00:002016-06-19 16:27:22updated18johnwaresjohn wares2016-06-19 05:03:34+00:002016-06-19 05:03:34updated17johnwaresjohn wares2016-06-19 00:19:10+00:002016-06-19 00:19:10updated16johnwaresjohn wares2016-06-18 19:37:20+00:002016-06-18 19:37:20updated15johnwaresjohn wares2016-06-18 18:56:54+00:002016-06-18 18:56:54updated14johnwaresjohn wares2016-06-18 16:39:15+00:002016-06-18 16:39:15updated13johnwaresjohn wares2016-06-18 15:55:16+00:002016-06-18 15:55:16updated12johnwaresjohn wares2016-06-18 10:37:56+00:002016-06-18 10:37:56updated11johnwaresjohn wares2016-06-17 05:16:48+00:002016-06-17 05:16:48updated10johnwaresjohn wares2016-06-17 05:16:30+00:002016-06-17 05:16:30updated9johnwaresjohn wares2016-06-17 05:15:18+00:002016-06-17 05:15:18updated8johnwaresjohn wares2016-06-17 05:11:21+00:002016-06-17 05:11:21updated7johnwaresjohn wares2016-06-17 05:09:25+00:002016-06-17 05:09:25updated6johnwaresjohn wares2016-06-16 19:53:48+00:002016-06-16 19:53:48updated5johnwaresjohn wares2016-06-16 19:08:22+00:002016-06-16 19:08:22updated4johnwaresjohn wares2016-06-16 19:07:35+00:002016-06-16 19:07:35updated3johnwaresjohn wares2016-06-16 19:05:26+00:002016-06-16 19:05:26updated2First createdjohnwaresjohn wares2016-06-16 18:37:09+00:002016-06-16 18:37:09created1wiki2016-06-30T15:22:33+00:00groups/eccofhl/wiki/ec337FalsePisaster Genotypes and Righting Response/groups/eccofhl/wiki/ec337/Pisaster_Genotypes_and_Righting_Response.htmljohn wares32 updatesPisaster Genotypes and Righting Response TL;DR Data! For this project, you will end up having 2-3 Pisaster ochraceus in your care; you must also keep your eyes on t...Falsejohnwares2016-06-30T15:22:33+00:00johnwaresjohn wares2016-06-29 15:51:46+00:002016-06-29 15:51:46updated15johnwaresjohn wares2016-06-29 04:26:27+00:002016-06-29 04:26:27updated14johnwaresjohn wares2016-06-28 04:28:38+00:002016-06-28 04:28:38updated13Added tag - hotjohnwaresjohn wares2016-06-24 15:41:01+00:002016-06-24 15:41:01addTag12johnwaresjohn wares2016-06-19 16:30:30+00:002016-06-19 16:30:30updated11johnwaresjohn wares2016-06-14 13:46:34+00:002016-06-14 13:46:34updated10johnwaresjohn wares2016-06-14 13:43:30+00:002016-06-14 13:43:30updated9Added tag - clinejohnwaresjohn wares2016-06-14 13:35:17+00:002016-06-14 13:35:17addTag8Added tag - phylodiversityjohnwaresjohn wares2016-06-14 13:35:16+00:002016-06-14 13:35:16addTag7Added tag - pcrjohnwaresjohn wares2016-06-14 13:35:08+00:002016-06-14 13:35:08addTag6Added tag - barnaclesjohnwaresjohn wares2016-06-14 13:35:07+00:002016-06-14 13:35:07addTag5johnwaresjohn wares2016-06-14 13:34:06+00:002016-06-14 13:34:06updated4johnwaresjohn wares2016-06-14 13:32:58+00:002016-06-14 13:32:58updated3johnwaresjohn wares2016-06-14 13:31:23+00:002016-06-14 13:31:23updated2First createdjohnwaresjohn wares2016-06-14 13:18:57+00:002016-06-14 13:18:57created1wiki2016-06-29T15:51:46+00:00groups/eccofhl/wiki/6f73aFalseBalanus diversity cline/groups/eccofhl/wiki/6f73a/Balanus_diversity_cline.htmljohn wares15 updatesBalanus diversity cline Background A few studies published DNA sequence data from barnacles (B. glandula) collected 15-20 years ago; the best example is here: ...Falsejohnwares2016-06-29T15:51:46+00:00Added tag - limpetsjohnwaresjohn wares2016-06-24 15:41:25+00:002016-06-24 15:41:25addTag10Added tag - diversityjohnwaresjohn wares2016-06-24 15:41:23+00:002016-06-24 15:41:23addTag9Added tag - distributionjohnwaresjohn wares2016-06-24 15:41:21+00:002016-06-24 15:41:21addTag8Added tag - ecologyjohnwaresjohn wares2016-06-24 15:41:20+00:002016-06-24 15:41:20addTag7Added tag - nichejohnwaresjohn wares2016-06-24 15:41:18+00:002016-06-24 15:41:18addTag6Added tag - hotjohnwaresjohn wares2016-06-24 15:41:17+00:002016-06-24 15:41:17addTag5johnwaresjohn wares2016-06-21 16:40:11+00:002016-06-21 16:40:11updated4morgankellyMorgan Kelly2016-06-20 14:33:01+00:002016-06-20 14:33:01updated3morgankellyMorgan Kelly2016-06-20 14:28:46+00:002016-06-20 14:28:46updated2First createdmorgankellyMorgan Kelly2016-06-20 14:11:36+00:002016-06-20 14:11:36created1wiki2016-06-21T16:40:11+00:00groups/eccofhl/wiki/d254bFalseTransect study/groups/eccofhl/wiki/d254b/Transect_study.htmljohn wares10 updatesTransect study In this exercise, you will use transect surveys and environmental data to create climate envelope models for a group of species of your choosing (it ...Falsejohnwares2016-06-21T16:40:11+00:00Added tag - hotmorgankellyMorgan Kelly2016-06-24 15:41:56+00:002016-06-24 15:41:56addTag5Added tag - adaptationmorgankellyMorgan Kelly2016-06-24 15:39:33+00:002016-06-24 15:39:33addTag4morgankellyMorgan Kelly2016-06-14 13:41:13+00:002016-06-14 13:41:13updated3morgankellyMorgan Kelly2016-06-14 01:37:59+00:002016-06-14 01:37:59updated2First createdmorgankellyMorgan Kelly2016-06-14 01:37:42+00:002016-06-14 01:37:42created1wiki2016-06-14T13:41:13+00:00groups/eccofhl/wiki/60339False1. Tigriopus heat tolerance/groups/eccofhl/wiki/60339/1_Tigriopus_heat_tolerance.htmlMorgan Kelly5 updates1. Tigriopus heat tolerance Background: Both genes and the environment contribute to most phenotypes. This complicates studies of local adaptation: local adaptation occurs when ...Falsemorgankelly2016-06-14T13:41:13+00:00hot/groups/eccofhl/search/index.rss?sort=modifiedDate&kind=all&sortDirection=reverse&excludePages=wiki/welcomelist/groups/eccofhl/search/?sort=modifiedDate&kind=all&sortDirection=reverse&excludePages=wiki/welcomeRecent ChangesRecentChangesListUpdates?sort=modifiedDate&kind=all&sortDirection=reverse&excludePages=wiki/welcome0/groups/eccofhl/sidebar/RecentChangesListmodifiedDateallRecent ChangesRecentChangesListUpdateswiki/welcomeNo recent changes.reverse5searchlist/groups/eccofhl/calendar/Upcoming EventsUpcomingEventsListEvents1Getting events…