Line 13: |
Line 13: |
| AG 501-X8 Resin (Mixed bed resin for deionizing formamide), 100g, BioRad #143-7424, $323 | | AG 501-X8 Resin (Mixed bed resin for deionizing formamide), 100g, BioRad #143-7424, $323 |
| | | |
− | [http://www.perkinelmer.com/Catalog/Product/ID/NEG034H250UC dATP-alphaS35], (250 uCi vial = 20 ul) [http://www.perkinelmer.com/AboutUs/ContactUs/ContactUS Perkin Elmer] #NEG034H250UC $172 | + | [https://www.revvity.com/product/datp-a-thio-35s-neg034h250uc dATP-alphaS35], (250 uCi vial = 20 ul) [https://www.perkinelmer.com/corporate/contactus Perkin Elmer] #NEG034H250UC $172 |
| :made first Tuesday of every month | | :made first Tuesday of every month |
| | | |
Line 105: |
Line 105: |
| '''DTT (dithiothreitol)''' | | '''DTT (dithiothreitol)''' |
| Aka Cleland's reagent; prevents oxidation of thiol groups and breaks down disulfide bonds. | | Aka Cleland's reagent; prevents oxidation of thiol groups and breaks down disulfide bonds. |
| + | |
| + | ==Buffer Amounts== |
| + | |
| + | * Denature salmon sperm by boiling for 10 minutes. |
| + | * Put DTT in foil-covered scintillation vial. |
| + | * Add SSC, formamide, TED, salmon sperm, & Denharts solution. |
| + | * Vortex. Keep heated at 55° C on heater block until use. |
| + | |
| + | <tab class="wikitable" sep="tab" border = "1" cellspacing="3" cellpadding = "3" head = top> |
| + | 10 ml - - - 8 ml - - - 5 ml - - - 4 ml - - - 3 ml - - - 2 ml - - |
| + | 150 mg DTT 120 mg DTT 75 mg DTT 60 mg DTT 45 mg DTT 30 mg DTT |
| + | 1.0 ml SSC 20x 800 ml SSC 20x 500 µl SSC 20x 400 µl SSC 20x 300 µl SSC 20x 200 µl SSC 20x |
| + | 6.0 ml Formamide 4.8 ml Formamide 3.0 ml Formamide 2.4 ml Formamide 1.8 ml Formamide 1.2 ml Formamide |
| + | 2.0 ml TED 5x 1.6 ml TED 5x 1 ml TED 5x 800 µl TED 5x 600 µl TED 5x 400 µl TED 5x |
| + | 1.6 ml Salmon Sperm 1.28 ml Salmon Sperm 800 µl Salmon Sperm 640 µl Salmon Sperm 480 µl Salmon Sperm 320 µl Salmon Sperm |
| + | 800 µl Denharts 50x 640 µl Denharts 50x 400 µl Denharts 50x 320 µl Denharts 50x 240 µl Denharts 50x 160 µl Denharts 50x |
| + | </tab> |
| | | |
| | | |