【心肌細胞總DNA】下面是細胞株培養的具體無菌技術要點：

1. 實驗開始前，無菌室和無菌操作臺需要紫外光照射30到60分鐘完全滅菌，用70%ethanol擦拭無菌操作臺面，并且打開無菌操作臺風扇運行10分鐘之后，才可以進行實驗操作。每次操作只能處理一株細胞株，即使培養基完全相同也不可共用培養基，以避免細胞間污染或失誤混淆。實驗完成后，請將實驗物品拿出工作臺，用70%ethanol再次擦拭無菌操作臺面。操作間隔應該讓無菌操作臺運轉10分鐘到15分鐘后，才能進行下一個細胞株的操作。

2. 無菌操作工作的區域應保持清潔和寬敞，必要物品（試管架、吸管、吸取器或吸管盒等）可以暫時放置，其它實驗用品使用完畢后應移出，有利于空氣流通。實驗用品需70%ethanol擦拭后方可帶入無菌操作臺內。實驗操作過程應在臺面的中央無菌區域內完成，請勿在邊緣非無菌區域進行操作。

3. 小心取用無菌實驗物品，避免細菌污染。請勿觸碰吸管尖頭部和容器瓶口處，也不要在打開的容器正上方進行操作。容器打開后，請用手夾住瓶蓋并輕握瓶身，傾斜大約45°角取用，盡量不要將瓶蓋口朝上放置于桌面。

4. 工作人員應當首先注意自身安全，必須穿戴齊實驗衣和實驗手套后才能進行實驗。對于病毒感染或是來自人類的細胞株應當特別小心操作，并選擇適當等級的無菌操作臺進行（至少是Class II）。操作過程中，應避免引起aerosol的產生，小心有毒性藥品，例如DMSO和TPA等，并避免針頭的傷害等等。

5. 定期檢測下列項目：

5.1. CO2鋼瓶的CO2壓力

5.2. CO2培養箱的CO2濃度、溫度、和水盤是否有污染（水盤的水需用無菌水，每周定時更換）。

5.3. 無菌操作臺內的airflow壓力，定期更換紫外線燈管和HEPA過濾膜，預濾網（300小時/預濾網，3000小時/HEPA）。

6. 水槽內可添加消毒劑（Zephrin 1:750），需定期更換水槽中的水。

【心肌細胞總DNA】齊一生物友情提醒，本產品僅供科研使用，不得用于臨床及診斷使用！

一細胞說明，具體詳細請見細胞附帶說明書

 二、客戶自備試劑

1、PBS  

2、 2、RPMI-1640 培養基（不含 Hepes）+ 10%胎牛血清+ 1%雙抗

3、0.25%  （W/V）胰蛋白酶（含 0.02% EDTA）

三、細胞背景

1、生長方式：貼壁

2、種屬：Homo sapiens  人

四、培養條件

1、培養基：RPMI-1640 培養基（不含 Hepes）+ 10%胎牛血清+ 1%雙抗

2、溫度：37.0°C

3、氣體:空氣  95%，CO2 5%

【心肌細胞總DNA】五、培養方法

 收到細胞后，在倒置顯微鏡下觀察整個細胞生長情況：

 如果細胞未長滿，用 75%酒精噴灑整個瓶消毒后放到超菌臺內，嚴格無菌操作，打開細胞培養瓶，留10ml 培養液繼續培養。如果細胞已長滿（達 80-90%）。即可進行傳代，具六、體步驟如下：

1，棄去培養液，用PBS洗 1-2次。

2，向瓶內加入 1.0-2.0ml 胰蛋白酶液，在倒置顯微鏡下觀察細胞消化情況，若細胞大部分變圓，迅速拿回操作臺，吸取胰蛋白酶，加含有 6ml  含 10%血清的培養液，輕輕吹打細胞。

3，加入等量的的培養液，輕輕吹打混勻后吸出一半，分到新的培養

4，傳代比例：1:2-1:3  

七、常見問題及解決方案

1、培養瓶有破裂，培養液有漏液：細胞極大可能會污染，所以我們會及時安排幫老師解決。

2、細胞漂�。号囵B瓶不開封，瓶口酒精擦拭后平躺放置在培養箱。次日觀察，如細胞大部分又貼回瓶底，表明細胞活力正常，剩余漂浮的細胞可以去掉，留 10ml 培養液培養觀察，細胞生長至匯合度 80%，進行消化傳代；如細胞還是不貼壁，將細胞離心收集轉到新培養瓶，原培養瓶加部分培養液繼續培養，中間注意觀察，我們的技術人員會一直跟蹤指導，直到問題解決。

細胞培養代理品牌

細胞培養代理品牌

【心肌細胞總DNA】齊一生物科技（上海）有限公司是一家專業生產和銷售各種生化試劑,細胞株、原代細胞、菌種、醫藥中間體,標準品和對照品的大型化學科技公司. 齊一生物銷售：021－6034 8496；181214 53965；173021 04490；Web:www.qiyibio.com

9236476  #Agreement, 9604, 2PM, 1 yr       An agreement for preventive maintenance provides coverage for travel, labor, ａnd parts, plus the cost of consumables required for two maintenance visits during 1 year. Work carried out during the preventive maintenance visit is fully documented in accordance with GMP/GLP directives.    CNY                                  

9232726  Block, 2-way, v1.02             CNY                                  

9239540  Cable Assy Extender, 2 wire, 22AWG, 12in          CNY                                  

9239396  Upgr. LiquiChip Analyzer (RUO)          CNY                                  

9239496  EX XY Motor Driver w/Encoder          CNY                            【心肌細胞總DNA】  

9239494  Sheath Fluid Tubing w/DeBubbler        CNY                                  

9239487  EX Analog PCBA           CNY                                  

9236515  Centrifuge, PM Service  "

Service response time (repair): not applicable

Periodic inspection/maintenance: yes

Inspection/maintenance delivery: onsite

Cost coverage for Repair Parts : not included

Cost coverage for labor: yes, full coverage

Cost coverage for travel: yes, full coverage

Replacement system (Loaner) provision: not applicable

Transportation cost coverage: yes, full coverage" CNY                                  

9232888  Camera, bar-code, BL180, P3xxx/BR3000          CNY                                  

9236636  Cover, Wash Station, BR8 RD              CNY                                  

9236451  Travel Charge, additional, 1 day           CNY                                  

9232725  Fitting, pumping tube, black         CNY                                  

9232712  Label, BioRobot 9604            CNY                                  

9236429  Sensor, plate switch, TWI            CNY                                  

9239142  Installation, BR MDx DSP            CNY                                  

9239070  Dilutor, Kloehn Versa3           CNY                                  

9232568  Power Cord, USA         CNY                                  

9238970  Target Prep, Premium Agreement  The Premium Cover Agreement TargetPrep provides full service coverage for the BioRobot TargetPrep. Costs for service visits are fully covered including travel, labor, ａnd parts. The agreement also includes one annual preventive maintenance visit, with costs for labor ａnd parts covered. With a Full Agreement, there is a priority response time of 24 hours (1 working day).     CNY                                  

9238791  Angle rotor for 8 Falcon tubes 50 ml           CNY                                  

9232328  Cable, power PCON98 board, BR8000        CNY                                  

9232188  T piece, pressure relief valve, dispense        CNY                            【心肌細胞總DNA】  

9238906  Upper casing, BS96              CNY                                  

9231863  Elution adapter, auto vac, BR8000        CNY                                  

9238681  Service manual Zent3, german             CNY                                  

9238680  Grease, load-bearing bolts     This is the grease for the Sigma centrifuge. Former cat no 2000770. CNY                                  

9231466  Spacer, DT-Adapter, BR8             CNY                                  

9014582  QIAVac Multiwell Base          CNY                                  

9022195  Reagent Chamber Lid, PyroQ24           CNY                                  

9022022  Sensor, Z-axis, with cable, QXA          CNY                                  

9021653  Kit, stickers, eluate drawer, Qsym        CNY                                  

9021623  Gateway 2, CC-Link, DCU           CNY                                  

9021289  Yellow grease for PM, EZ1           CNY                                  

9240671  Reservoir, 500ml w/cap, AP96/196              CNY                                  

QY-x518  SK-BR-3人乳腺癌細胞                                   

QY-x519  SK-MEL-1人皮膚黑色素瘤細胞                                    【心肌細胞總DNA】

QY-x520  SK-N-SH人神經母細胞瘤                                     

QY-x521  SK-OV-3人卵巢腺瘤細胞                                      

QY-x522  SMMC-7721人肝癌細胞                                       

QY-x523  SW13人腎上腺皮質瘤                                   

QY-x524  SW480人結直腸癌                                        

QY-x525  T84人結腸癌細胞                                   

QY-x526  THP1人單核細胞型淋巴瘤                                    

QY-x527  U-2 OS人骨肉瘤細胞                                     

QY-x528  U251人神經膠質細胞瘤                                        

QY-x529  U-937人淋巴瘤細胞                                       

QY-x530  3T3 swissswiss鼠胚胎成纖維細胞                                       

QY-x531  3T3L1小鼠胚胎成纖維細胞(前脂肪)                                   

QY-x532  3T6swiss小鼠胚胎成纖維細胞                                     

QY-x533  BA/F3小鼠原B細胞                                      

QY-x534  BHK-21金黃地鼠腎                                        

QY-x535  BS-C-1非注洲綠猴腎                                      【心肌細胞總DNA】

QY-x536  C2C12小鼠成肌細胞                                      

QY-x537  C3H 10T1/2 2A6小鼠成纖維細胞                                         

QY-x538  CHOdhfr二氫葉酸缺陷型中國倉鼠卵巢細胞                                      

QY-x539  CHO-K1中國倉鼠卵巢細胞                                   

QY-x540  COS-1非洲綠腎                                      

QY-x541  COS-7非洲綠猴腎細胞                                         

QY-x542  CV-1猴腎細胞                                        

QY-x543  IAR20小鼠肝細胞                                   

QY-x544  IEC-6大鼠小腸隱窩上皮細胞                                       

QY-x545  LL-PK1,豬腎細胞                                    

QY-x546  MC3T3-E1小鼠胚胎成骨細胞                                      

QY-x547  MDCK狗腎細胞                                     

QY-x548  MEF小鼠胚胎成纖維細胞