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==Our Lab's Knowledge Base==
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#redirect[[Lab Knowledge Base]]
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==Other optional things to do==
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#On this page: [[image_logo_url]] put just the url for the logo that you want in the upper left corner of your wiki.
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*Control the rights of anonymous and logged in users through the [[Control Panel]] (which can be found in the toolbox on the left side of this page).
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*Add the url for your wiki to the [http://www.editthis.info/wiki/index.php/Categorized_Wiki_List Categorized wiki list]. Simply edit that page and put your wiki under the appropriate category.
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*To increase the amount of space you have for uploads, add links to http://editthis.info from external sites, and go to the [http://editthis.info/wiki/index.php/Link_Quota_Page Link Quota Page] to increase your uploads quota.
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*Edit [[MediaWiki:Sidebar]] to change the navigation menu
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*Add this wiki to some external wiki indexes:
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#[http://en.wikipedia.org/wiki/List_of_wiki Wikipedia list of wikis]
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#[http://www.wikiindex.com/Add_a_Wiki WikiIndex]<br>
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*Read the [http://meta.wikipedia.org/wiki/MediaWiki_User%27s_Guide User's Guide] for other usage and configuration help.
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<pre>AF-6 Medium
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Note:  All nutrient stock solutions are stored at 4˚C in the walk-in and the trace metal and vitamin stocks are in the -20˚C (thaw these completely in warm water bath with parafilm around caps or at room temperature…invert several times before using).
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1.  Begin with 600mL of distilled water in the plastic 1L graduated cylinder.  Continue the rest of the process in the sterile environment of the transfer hood.
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2.  Add 10mL of each 100X Nutrient Stock Solutions to the cylinder (using different sterile volumetric pipets for each solution).
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Nutrient Stocks AF-6 Medium 100X Stock solutions
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#1 NaNO3 14.00 mg 1.40 g/100mL
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#2 NH4NO3 2.20 mg 0.22 g/100mL
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#3 MgSO4 x 7H20 3.00 mg 0.30 g/100mL
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#4 KH2PO4 1.00 mg 0.10 g/100mL
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#5 CaCl2 x H20 1.00 mg 0.10 g/100mL
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#6 K2HPO4 0.50 mg 0.05 g/100mL
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#7 Fe-citrate 0.20 mg 0.02 g/100mL
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#8 Citric acid 0.20 mg 0.02 g/100mL
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#9 MES buffer* 40.00 mg 4.00 g/100mL
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*MES = 2-morpholinoethanesulfonic acid monohydrate 98% from VWR (cat.# AAA16104-22)
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_____________________________________________________________________________
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3.  Add 5mL of the P IV Trace Metal Working Solution.
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P IV Trace Metal Working Solution:  Start with 85mL of distilled water.  Add 0.1 g of Na2EDTA x 2H2O, stirring continuously.  Add 1mL of each of the FIVE trace metal 100X stock solutions.  Bring final volume to 100mL with distilled water.
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P IV Trace Metals Working Solution 100X Stock Solution
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FeCl3 x 6H20   19.60 mg 1.960 g/100mL
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MnCl2 x 4H20     3.60 mg 0.360 g/100mL
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ZnSO4 x 7H20     2.20 mg 0.220 g/100mL
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CoCl x H20     0.40 mg 0.040 g/100mL
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Na2MoO4 x 2H20     0.25 mg 0.025 g/100mL
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Na2EDTA x 2H20 100.00 mg 0.100 g crystals directly
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_____________________________________________________________________________
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4.  Add 10uL of each of the Vitamin Stocks.
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Vitamin Stocks AF-6 Medium 100,000X Stock Solutions
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Biotin (vitamin H or B7) 0.2 ug 0.02 g/100mL†
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Thiamine HCl (vitamin B1) 1.0 ug 0.10 g/  10mL
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Vitamin B6 (pyridoxine) 0.1 ug 0.01 g/  10mL
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Vitamin B12 (cyanocobalamin) 0.1 ug 0.01 g/  10mL‡
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† Instead of 10uL of 0.02g/100mL stock solution, can add 20uL of 0.1g/L stock solution.
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‡ Instead of 10uL of 0.01g/ 10mL stock solution, can add 10uL of 1.0g/L stock solution.
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_____________________________________________________________________________
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5.  Add 7.5mL of undiluted Carolina Biological Supply Soil Water Supernatant for each liter of AF-6.  If diluted to the specified 50mL of Soil Water Supernatant into 1L of distilled water, then add 150mL to the AF-6 cylinder mix instead.
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6.  The solution will be acidic, adjust pH to 6.6 with 1N NaOH (=1M; found in chemical cabinet, use NaOH hydroxide pellets to make more solution if necessary).  Use pH paper or pH meter.  If overshot, then add a drop or two of concentrated HCl (acid storage under fume hood).
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7.  Bring final volume up to 1000mL with distilled water.
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8.  Aliquot media into media bottles and autoclave and then allow to cool before using.
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<pre>
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'''Bold text''''''Bold text'''
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</pre>
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Current revision as of 16:20, 20 May 2007

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