訂做你的阿凡達：光磁控遠端神經操作
(Tailoring Your Avatar
--Optomagnetogenetic Tele-Neurocontrol)

AMB-1（日本教授2001 paper裡面用的。Effects of growth medium composition, iron sources and atmospheric oxygen concentrations on production of luciferase-bacterial magnetic particle complex by a recombinant Magnetospirillum magneticum AMB-1）Medium Receipt(protocol):1 liter of magnetic spirillum growth medium (MSGM) contains…. Distilled water 1.0 L 10 ml of Wolfe’s vitamin solution 5 ml of Wolfe’s mineral solution 2 ml of ferric quinate solution (0.27 g of FeCl3 and 0.19 g of quinic acid in 100 ml water) 0.68 g of KH2PO4  0.12 g of sodium nitrate NaNO3 0.74g of succinic acid  0.1 g of sodium thioglycolate＊ Enriched MSGM contains 0.02% polypeptone 0.01% yeast extract 0.005% L-cysteine instead of sodium thioglycolate.  All of the media used were supplemented with 5 mg/ml tetracycline pH 6.75 with aqueous sodium hydroxide. Microaerobic ＊ 0.01 M Ferric Quinate: FeCl3 .....................................0.27 g Quinic Acid (Sigma Q-0500)................0.19 g Distilled water...........................100.0 ml Dissolve and autoclave at 121C for 15 minutes. ＊ Wolfe's Vitamin Solution: Available from ATCC as a sterile ready-to-use liquid (Vitamin Supplement, catalog no. MD-VS).Biotin....................................2.0 mg Folic acid................................2.0 mg Pyridoxine hydrochloride..................10.0 mg Thiamine . HCl............................5.0 mg Riboflavin................................5.0 mg Nicotinic acid............................5.0 mg Calcium D-(+)-pantothenate................5.0 mg Vitamin B12...............................0.1 mg p-Aminobenzoic acid.......................5.0 mg Thioctic acid.............................5.0 mg 2Distilled water...........................1.0 L 3* Wolfe's Mineral Solution: Available from ATCC as a sterile ready-to-use liquid (Trace Mineral Supplement, catalog no. MD-TMS.)Nitrilotriacetic acid.....................1.5 g MgSO4 . 7H2O ..............................3.0 g MnSO4 . H2O ...............................0.5 g NaCl......................................1.0 g FeSO4 . 7H2O ..............................0.1 g CoCl2 . 6H2O ..............................0.1 g CaCl2 .....................................0.1 g ZnSO4 . 7H2O ..............................0.1 g CuSO4 . 5H2O ..............................0.01 g AlK(SO4)2 . 12H2O..........................0.01 g H3BO3 .....................................0.01 g Na2MoO4 . 2H2O.............................0.01 g Distilled water...........................1.0 L Add nitrilotriacetic acid to approximately 500 ml of water and adjust to pH 6.5 with KOH to dissolve the compound. Bring volume to 1.0 L with remaining water and add remaining compounds one at a time.

Still aiting the author of the 中正大學's paper entitled"2009磁鐵細菌（地球磁場的領航員" to share his protocol in this paper with us, who is the 畢業研究生 潘信凱。And we had asked the director professor of this paper in the 中正大學, he promised to sent the strain MS-1 in two days(5/31-6/2) So right after we receive the bac MS-1 and the protocol from 學長, we'll start to culture MS-1 immediately.

以下是目前最新進度0531，隨時更新！要菌的進度（因為採買上的困難，先以跟別的實驗室要為主）：我們目前最想要的菌是AMB-1（日本教授2001 paper裡面用的。Effects of growth medium composition, iron sources and atmospheric oxygen concentrations on production of luciferase-bacterial magnetic particle complex by a recombinant Magnetospirillum magneticum AMB-1），但於兩週前就已寫信兩三次給日本教授跟他們要AMB-1，但都沒回信仍石沈大海中QQ所以我們決定跟中正大學的陳建易教授要菌（2009磁鐵細菌（地球磁場的領航員）但他們用的是類似的strain──MS-1。但因為這篇paper還在保護期不公開內容，很久前我便跟這篇碩士論文的學長聯絡，學長表示這兩天會寄給我paper。而五天前陳教授也很熱心的打電話給我，說可以給我們MS-1，也會幫我們寫信給德國大學要AMB-1！陳教授請他的助理寄了，這兩天應該會收到MS-1。配置Medium（MSGM）的進度：我們預定使用AMB-1，所以我們的protocol參考日本教授2001那篇為主，但是iGEM實驗室現在還缺滿多需要的藥品，學長建議我們直接去買現成的medium，並推薦我們以下兩種：ATCC medium1653 http://www.atcc.org/Attachments/2856.pdfBCRC medium 610 https://www.bcrc.firdi.org.tw/BSAS_cart/controller?event=SEARCH&bcrc_no=17316&type_id=3&keyword=Magnetospirillum內容物看起來日本教授的protocol和ATCC（國外廠商）賣的比較像，但運送時間會比較久起碼兩週以上。而BCRC（台灣）賣的和目前我們的protocol相較則是缺滿多東西，但缺的東西比較好找，可能就我們買了自己再加進去。其實也不用堅持一定要和日本那些不回信的教授（= = +）的protocol一樣，因為那篇是改良的protocol，照著做只是可以提昇菌或BMP的產率而已。since我們的目標不是養出超級多的磁菌和BMP，我們只要菌養的出來、菌又生的出BMP、BMP又有活性這樣就夠了。而市面上現成的medium和protocol差別只在於一些營養成分和iron source的不同，學長也說菌種類似，medium只要差不多其實都養的出來。我們也可以在養的過程中做修正與改良。但有鑑於目前AB組都急迫的要開始養菌，所以我們先拿這兩天會到的中正MS-1來養，protocol也先參考中正大學那篇，或者購買BCRC的medium再做修正。所以最快一個禮拜內就可以開始養菌了，而菌在日本教授那篇裡面最理想的狀況需要4天長成（有關鍵的L-cystein情況下, which we and the 現成medium don’t have@@）。所以最快最快十天，六月中左右AB組就可以開始進行實驗了。給C’的組員：如果上面有任何不懂的……快回去做功課！！ =”= 讀paper！至少讀我等等寄給大家那篇日本大學 AMB-1的protocol裡我劃的重點，非常簡單！看過就會對medium的protocol有概念了。

Medium Receipt(protocol):1 liter of magnetic spirillum growth medium (MSGM) contains…. Distilled water 1.0 L 10 ml of Wolfe’s vitamin solution 5 ml of Wolfe’s mineral solution 2 ml of ferric quinate solution (0.27 g of FeCl3 and 0.19 g of quinic acid in 100 ml water) 0.68 g of KH2PO4  0.12 g of sodium nitrate NaNO3 0.74g of succinic acid  0.1 g of sodium thioglycolate＊ Enriched MSGM contains 0.02% polypeptone 0.01% yeast extract 0.005% L-cysteine instead of sodium thioglycolate.  All of the media used were supplemented with 5 mg/ml tetracycline pH 6.75 with aqueous sodium hydroxide. Microaerobic ＊ 0.01 M Ferric Quinate: FeCl3 .....................................0.27 g Quinic Acid (Sigma Q-0500)................0.19 g Distilled water...........................100.0 ml Dissolve and autoclave at 121C for 15 minutes. ＊ Wolfe's Vitamin Solution: Available from ATCC as a sterile ready-to-use liquid (Vitamin Supplement, catalog no. MD-VS).Biotin....................................2.0 mg Folic acid................................2.0 mg Pyridoxine hydrochloride..................10.0 mg Thiamine . HCl............................5.0 mg Riboflavin................................5.0 mg Nicotinic acid............................5.0 mg Calcium D-(+)-pantothenate................5.0 mg Vitamin B12...............................0.1 mg p-Aminobenzoic acid.......................5.0 mg Thioctic acid.............................5.0 mg 2Distilled water...........................1.0 L 3* Wolfe's Mineral Solution: Available from ATCC as a sterile ready-to-use liquid (Trace Mineral Supplement, catalog no. MD-TMS.)Nitrilotriacetic acid.....................1.5 g MgSO4 . 7H2O ..............................3.0 g MnSO4 . H2O ...............................0.5 g NaCl......................................1.0 g FeSO4 . 7H2O ..............................0.1 g CoCl2 . 6H2O ..............................0.1 g CaCl2 .....................................0.1 g ZnSO4 . 7H2O ..............................0.1 g CuSO4 . 5H2O ..............................0.01 g AlK(SO4)2 . 12H2O..........................0.01 g H3BO3 .....................................0.01 g Na2MoO4 . 2H2O.............................0.01 g Distilled water...........................1.0 L Add nitrilotriacetic acid to approximately 500 ml of water and adjust to pH 6.5 with KOH to dissolve the compound. Bring volume to 1.0 L with remaining water and add remaining compounds one at a time.

老師滿希望能著重這部分，回去想想這也是舉辦human practice的原意嘛>///<！