tag:blogger.com,1999:blog-244942332011-06-17T20:06:10.073-07:00DNA LabTrininoreply@blogger.comBlogger71100tag:blogger.com,1999:blog-24494233.post-1142981722482947082006-03-21T14:36:00.000-08:002006-03-21T14:55:22.493-08:00DNA Extraction NotesDid small test using supernatant from centrifugation step with K 1 (in Koba Extraction) samples #62, 63 (Sheet BH)Results: Got DNA pellet.Need to check to see what yield looks like and how it works for PCR.Trininoreply@blogger.com0tag:blogger.com,1999:blog-24494233.post-1142981825705392612006-03-08T14:55:00.000-08:002006-03-21T14:57:05.706-08:00Prep, pptn etcPrepared leaf tissue (BG 1-36) for DNA extraction tomorrow.Autoclaved tubes, tips for extraction.Precipitated DNA for extracts BF 37-72: Samples BF 47, 55 and 71 had very tiny pellets prior to centrifugation.Trininoreply@blogger.com0tag:blogger.com,1999:blog-24494233.post-1142981964164667652006-03-07T16:58:00.000-08:002006-03-21T14:59:24.163-08:00DNA Extraction etcSheet BF 37-72 last fp#: 3336Leaf tissue for extraction prepared on previous day.Changed waterbath waterGave MB list of items to order.Prepared tips & tubes for autoclaving.Trininoreply@blogger.com0tag:blogger.com,1999:blog-24494233.post-1142981998935587832006-03-06T17:59:00.000-08:002006-03-21T15:00:20.596-08:00QuestionsHow long does 0.5M EDTA (pH 8.0) last on benchtop/in fridge?Other chemicals whose expiration/half-life is unknown: Sorbitol (solid and 1.75M prep), CTAB, N-Lauroyl Sarcosine.Trininoreply@blogger.com0tag:blogger.com,1999:blog-24494233.post-1142982234818673722006-02-10T15:01:00.000-08:002006-03-21T15:03:54.820-08:00AboutA CRU DNA Lab chronicle of observations, notes, protocols, recipes etc and other general information which maybe useful to archive.Trininoreply@blogger.com0tag:blogger.com,1999:blog-24494233.post-1142982443960414032006-01-21T15:07:00.000-08:002006-03-21T15:09:48.000-08:00DNA ExtractionsChemicals used (approximation)70 mL CIAA per 100 extracts100 mL Isopropanol per 100 extracts100 mL Ethanol per 100 extractsNotesD-squared note: Overnight pptn of DNA ok. Pellets unaffected.EDTA Used in March 2002: Disodium dihydrateGel quantitation: Dryloading works well only if yield is good.Trininoreply@blogger.com0tag:blogger.com,1999:blog-24494233.post-1142982400745505932006-01-21T15:06:00.000-08:002006-03-21T15:09:16.340-08:00PCR and ElectrophoresisPCR NotesTAQ + Buffer + Magnesium Chloride 50U/mL: 0.1 mL per reactionGroup A: 27/28; 45A/45B; 53/54B; 132/133Group B?: 240/241; 246/247Group C: 86/87Group D: 35/36; 43/44Electrophoresis NotesLadder (Lambda) 114ng/mL: 25 mL per gelLong Ranger gelsSigma Urea dissolves inTrininoreply@blogger.com0