植物组织切片及原位杂合反应ISH-中央研究院南部生物技术中心

植物组织切片及原位杂合反应ISH-中央研究院南部生物技术中心 單A 中央研究院南部生物技術中心 植物組織切片及原位雜合反應(ISH)服務申請表 Plant Tissue Section and ISH Service Application Form 中華民國 年 月 日(填表日期) 實驗室主持人 PI, 電話Tel., E-mail: 委託者姓名, 電話 Tel., E-mail: Applicant 【請委託者先詳讀注意事項】 提醒,每個實驗室切片樣本送件,一個月以四種樣本為上限;ISH送件,限同種樣本,二個月以四種基因為上限。 For each Lab., each month can send maximum 4 tissue sectioning samples and each two months maximum 4 genes for ISH service. A._____________ 植物種類 Plant species, 1)蝴蝶蘭Phalaenopsis orchid; 2)水稻 Rice; 3)阿拉伯芥Arabidopsis; 4)其他(另議) Other (need to be discussed),_____________ B._____________ 需服務的項目Service item,1) Tissue sample collection & fixation, 2) Tissue sectioning, 3) In situ hybridization, 4) Take microscopy photograph 1. 組織切片 Tissue sectioning: Each sample=_____ slides, Thickness of tissue section= _____µm aSample No. 1.1樣本組1.2組織切1.4組織化學1.5目標基1.3組織切備註 b織和數量 面(橫切/縱切) 片特殊切點染色 (請參閱因名稱或代Remarks 位置 (ex.花以下說明) 號(如不需ISHTissue Cross/ 苞的生長點) Histochemical 服務可略過) sample & vertical bstaining section quantity Special organ ISH target gene/ Code 1 1 2 3 4 Notes, a 樣本組織,數量, Tissue sample (quantity), a)根Root ; b)葉Leaf ; c)花苞Floral bud ; d)圓球體protocorm ; e) 擬圓球體PLB ; f)其他Other, b 組織化學染色Histochemical staining: 1) TBO; 2) Hematoxylin; 3) DAPI; 4) Other: __________ 注意: 若進行ISH則不在同一玻片另行組織染色,但可另外切片單獨作組織染色。 *切片完成後不進行ISH,則不需填以下表格。 *If not proceed ISH service, no need to fill the following table. 2. ISH information資訊: (請確認後勾選 ,) RNA Probe Probe No. 目標基因名要委託要顯微組織內備註 probe size is high Remarks dot (bp) (自mRNAgene 稱或代號 合成探照相ISH行合成detection specific表現量probe需填conc. (自Name of 針 基因專結果行合成probe寫此欄) target gene/ (已進行強需填寫此欄) Code To take ISH Request 一性高 RT/ qRT- PCRphotograph probe 確認). (自行合成synthesis Gene probe需填expression 寫此欄) level 1 2 2 3 4 請簡單描述目標基因之功能、基因表達之組織部位和時期(請附件qRT-PCR實驗結果)、及參考文獻. Please descript your target gene’s function, tissue localization & expression stage (Kindly attach qRT-PCR data). reference of ISH picture similar to your gene or in other plant is preferred [in Pdf file]. 注意事項: 1. 送件之前,請填妥此服務申請表( 本表須詳實填寫,未依規定填寫之處,概由申請人自行負責 )。 2. 若申請者欲自行採樣,每個樣本提供10個以上組織塊 (可能因測試難度而異),送件切片厚度 以3-5 mm為主,請以鋒利乾淨刮鬍刀或解剖刀新鮮採樣,並置於新鮮配置固定液(成分, 4%paraformadehyde +0.1%Tween 20 +0.1%TritonX-100+ 0.25%glutaldehyde/ 1XPBS),4?抽氣 (680~720 mmHg) overnight,隔日將樣本取出並以1xPBS/ DEPC-HO 清洗1次(10min),置冰上2 依序每隔30min換置30%、50%最後放在70%酒精/ DEPC-HO,低溫(冷藏)送至本實驗室。若2 由本實驗室取樣,請確實說明採樣的位置及材料有無特殊採樣時期的需求。 3. 請事先告知原位雜合反應完成後是否封片或特殊染色,否則以一律封片為原則。拍照檔案以tif 檔儲存,並於完成後將拍照檔案交與申請人。本實驗室僅提供植物組織原位雜合反應服務,不 負責基因表現結果之解讀。 4. 服務連絡人,連宜珍 小姐,電話06-5056630#201,E-mail: tokyla09111@gmail.com NOTICE: 3 1. Before sending sample, please complete the application form for this service (This form should be filled correctly, if not, the applicant is solely responsible for its own). 2. If applicants wish to collect sample themselves, please provide each sample >10 tissue blocks with thickness of 3-5 mm. Please use clean and sharp razor to collect fresh sample and place in fresh PFA fixation solution (4%paraformadehyde +0.1%Tween 20 +0.1%TritonX-100+ 0.25%glutaldehyde/ DEPC-HO), exhaust samples 2owith 680~720 mmHg at 4C overnight. Then wash 10 min. with 1xPBS/ DEPC-HO, serial dehydration with 2 30%EtOH, 50%EtOH, and keep in 70%EtOH (in DEPC treated water), and then cold send to Lab201. If need us to collect sample, please inform us the location and date of sampling. 3. After in situ hybridization, slides will be mounted, if you need special chemical staining, please inform us in advance. Microscopy photos are saved in Tif files and photo files upon completion will delivery to the applicant. We provides only plant tissue in situ hybridization reactions services and not responsible for the interpretation of the results. 4. Service contact person: Ms. Yi-Chen Lien Tel: 06-5056630#201,E-mail: tokyla09111@gmail.com. 實驗室主持 人簽名 ?本人確已詳閱送件要求說明, Signature of PI 並同意所載注意事項之約定。 委託者 I have read and agreed to follow the instructions 簽 名 Signature of applicant 以下由本實驗室填寫,This part filled by our Lab. 收件日期Date received 年 月 日 服務編號 Year Month Day Service No. 核 定 人 Approved by 服務完成人 交 件 簽 名 日 期 Service completed by Completion date 4