【心肌细胞总DNA】下面是细胞株培养的具体无菌技术要点：

1. 实验开始前，无菌室和无菌操作台需要紫外光照射30到60分钟完全灭菌，用70%ethanol擦拭无菌操作台面，并且打开无菌操作台风扇运行10分钟之后，才可以进行实验操作。每次操作只能处理一株细胞株，即使培养基完全相同也不可共用培养基，以避免细胞间污染或失误混淆。实验完成后，请将实验物品拿出工作台，用70%ethanol再次擦拭无菌操作台面。操作间隔应该让无菌操作台运转10分钟到15分钟后，才能进行下一个细胞株的操作。

2. 无菌操作工作的区域应保持清洁和宽敞，必要物品（试管架、吸管、吸取器或吸管盒等）可以暂时放置，其它实验用品使用完毕后应移出，有利于空气流通。实验用品需70%ethanol擦拭后方可带入无菌操作台内。实验操作过程应在台面的中央无菌区域内完成，请勿在边缘非无菌区域进行操作。

3. 小心取用无菌实验物品，避免细菌污染。请勿触碰吸管尖头部和容器瓶口处，也不要在打开的容器正上方进行操作。容器打开后，请用手夹住瓶盖并轻握瓶身，倾斜大约45°角取用，尽量不要将瓶盖口朝上放置于桌面。

4. 工作人员应当首先注意自身安全，必须穿戴齐实验衣和实验手套后才能进行实验。对于病毒感染或是来自人类的细胞株应当特别小心操作，并选择适当等级的无菌操作台进行（至少是Class II）。操作过程中，应避免引起aerosol的产生，小心有毒性药品，例如DMSO和TPA等，并避免针头的伤害等等。

5. 定期检测下列项目：

5.1. CO2钢瓶的CO2压力

5.2. CO2培养箱的CO2浓度、温度、和水盘是否有污染（水盘的水需用无菌水，每周定时更换）。

5.3. 无菌操作台内的airflow压力，定期更换紫外线灯管和HEPA过滤膜，预滤网（300小时/预滤网，3000小时/HEPA）。

6. 水槽内可添加消毒剂（Zephrin 1:750），需定期更换水槽中的水。

【心肌细胞总DNA】齐一生物友情提醒，本产品仅供科研使用，不得用于临床及诊断使用！

一细胞说明，具体详细请见细胞附带说明书

 二、客户自备试剂

1、PBS  

2、 2、RPMI-1640 培养基（不含 Hepes）+ 10%胎牛血清+ 1%双抗

3、0.25%  （W/V）胰蛋白酶（含 0.02% EDTA）

三、细胞背景

1、生长方式：贴壁

2、种属：Homo sapiens  人

四、培养条件

1、培养基：RPMI-1640 培养基（不含 Hepes）+ 10%胎牛血清+ 1%双抗

2、温度：37.0°C

3、气体:空气  95%，CO2 5%

【心肌细胞总DNA】五、培养方法

 收到细胞后，在倒置显微镜下观察整个细胞生长情况：

 如果细胞未长满，用 75%酒精喷洒整个瓶消毒后放到超菌台内，严格无菌操作，打开细胞培养瓶，留10ml 培养液继续培养。如果细胞已长满（达 80-90%）。即可进行传代，具六、体步骤如下：

1，弃去培养液，用PBS洗 1-2次。

2，向瓶内加入 1.0-2.0ml 胰蛋白酶液，在倒置显微镜下观察细胞消化情况，若细胞大部分变圆，迅速拿回操作台，吸取胰蛋白酶，加含有 6ml  含 10%血清的培养液，轻轻吹打细胞。

3，加入等量的的培养液，轻轻吹打混匀后吸出一半，分到新的培养

4，传代比例：1:2-1:3  

七、常见问题及解决方案

1、培养瓶有破裂，培养液有漏液：细胞极大可能会污染，所以我们会及时安排帮老师解决。

2、细胞漂浮：培养瓶不开封，瓶口酒精擦拭后平躺放置在培养箱。次日观察，如细胞大部分又贴回瓶底，表明细胞活力正常，剩余漂浮的细胞可以去掉，留 10ml 培养液培养观察，细胞生长至汇合度 80%，进行消化传代；如细胞还是不贴壁，将细胞离心收集转到新培养瓶，原培养瓶加部分培养液继续培养，中间注意观察，我们的技术人员会一直跟踪指导，直到问题解决。

细胞培养代理品牌

细胞培养代理品牌

【心肌细胞总DNA】齐一生物科技（上海）有限公司是一家专业生产和销售各种生化试剂,细胞株、原代细胞、菌种、医药中间体,标准品和对照品的大型化学科技公司. 齐一生物销售：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                            【心肌细胞总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                            【心肌细胞总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人乳腺癌细胞                                   

QY-x519  SK-MEL-1人皮肤黑色素瘤细胞                                    【心肌细胞总DNA】

QY-x520  SK-N-SH人神经母细胞瘤                                     

QY-x521  SK-OV-3人卵巢腺瘤细胞                                      

QY-x522  SMMC-7721人肝癌细胞                                       

QY-x523  SW13人肾上腺皮质瘤                                   

QY-x524  SW480人结直肠癌                                        

QY-x525  T84人结肠癌细胞                                   

QY-x526  THP1人单核细胞型淋巴瘤                                    

QY-x527  U-2 OS人骨肉瘤细胞                                     

QY-x528  U251人神经胶质细胞瘤                                        

QY-x529  U-937人淋巴瘤细胞                                       

QY-x530  3T3 swissswiss鼠胚胎成纤维细胞                                       

QY-x531  3T3L1小鼠胚胎成纤维细胞(前脂肪)                                   

QY-x532  3T6swiss小鼠胚胎成纤维细胞                                     

QY-x533  BA/F3小鼠原B细胞                                      

QY-x534  BHK-21金黄地鼠肾                                        

QY-x535  BS-C-1非注洲绿猴肾                                      【心肌细胞总DNA】

QY-x536  C2C12小鼠成肌细胞                                      

QY-x537  C3H 10T1/2 2A6小鼠成纤维细胞                                         

QY-x538  CHOdhfr二氢叶酸缺陷型中国仓鼠卵巢细胞                                      

QY-x539  CHO-K1中国仓鼠卵巢细胞                                   

QY-x540  COS-1非洲绿肾                                      

QY-x541  COS-7非洲绿猴肾细胞                                         

QY-x542  CV-1猴肾细胞                                        

QY-x543  IAR20小鼠肝细胞                                   

QY-x544  IEC-6大鼠小肠隐窝上皮细胞                                       

QY-x545  LL-PK1,猪肾细胞                                    

QY-x546  MC3T3-E1小鼠胚胎成骨细胞                                      

QY-x547  MDCK狗肾细胞                                     

QY-x548  MEF小鼠胚胎成纤维细胞