MICRO-ePUMP Microinjector with Cell Penetrator
- INTRODUCTORY PRICE FOR A LIMITED TIME
Regulated compensation and injection pressure
Pressure output: 0.3–87 PSI
Onboard MICRO-ePORE™
Pinpoint Cell Penetrator for targeted microinjection and increased viability of injected embryos
Audio continuity tone indicating active probe
Injection counter to indicate total number of injections
User-adjustable frequency and voltage through touch screen
Foot switch to activate voltage for MICRO-ePORE™ and injection
Low volume tubing assembly
Benefits
- Internal pressure source
- Intuitive user-interface for injectorand MICRO-ePORE™
- Small footprint takes up very littlebench space
- Easy to navigate with touch screenand control knob
- Inject into a single cell inpicoliter volumes
Applications
- Microinjection of diverse compounds and biomolecules – DNA, RNA, proteins
- Pre- and post-implantation in embryos of various species – mice, rats, monkeys, bovine, pigs, zebrafish, etc.
Designed to simplify intracellular injection and a variety of other microinjection tasks, the MICRO-ePUMP uses carefully regulated air pressure for injecting cells with fluid. Injected volumes range from picoliters to nanoliters. The port supplies positive pressure for high-pressure ejection. The pressure port maintains a low positive “compensation” pressure to the injecting pipette between injection pulses to prevent fluid uptake through capillary action.
Timing, injection pressure and compensation pressure are adjusted independently using the touch screen interface. Time intervals can range from 2 seconds down to 10 ms or less, depending on the injection pressure setting. The injection pressure interval is triggered using the foot switch.
The MICRO-ePUMP is designed to inject very small quantities of fluids, such as drugs, into cells or small organelles. Pressure injection is an especially useful alternative to electroionophoresis, since it does not mandate the use of charged ions. The compensation pressure is a constant low pressure that eliminates any capillary action front-filling of the pipette and then the precise burst of regulated higher pressure is activated with the foot switch that is included.
WPI’s MICRO-ePORE™ Pinpoint Cell Penetrator technology is embedded inside the MICRO-ePUMP. When the researcher enables the MICRO-ePORE™, it delivers a highly localized voltage signal to a targeted injection site to facilitate penetration with minimal trauma. The researcher determines the amplitude and frequency of the signal that best suits the application. The signal originates in the control box, and it is transmitted through the electrode interface cable to the microelectrode holder. A silver wire is used to transmit the signal into the electrically conductive substance being injected. A reference electrode is used to place the media at 0.0 V potential with reference to the generated voltage.
The MICRO-ePUMP offers separate regulated compensation (back filling prevention) and injection pressures with a precision timing circuit that switches from injection pressure to compensation pressure automatically.
System components
What is included | Qty |
MICRO-ePUMP | 1 |
300863 WPI MICRO-ePORE™ Electrode Holders(Electrode holder includes an electrode holderstartup kit.) | 2 |
Microelectrode Holder Interface Cable | 1 |
Well-style Reference Electrode | 1 |
Probe-style Reference Electrode Assembly | 1 |
5430-ALL PicoNozzle Kit which includes:(2) PicoNozzle tip assemblies(Handle diameter is 6.25 x 100 mm.)(2) 5’ tubing(4) 1.0 mm pipette gaskets (green)(4) 1.2 mm pipette gaskets (black)(4) 1.5 mm pipette gaskets (red)(4) 1.65 mm pipette gaskets (white) | 1 |
(1) Foot switch | 1 |
(1) AC/DC 24 V Power Adapter | 1 |
(1) Instruction Manual (available online at www.wpiinc.com/manuals) | 1 |
Microinjection System
MICRO-ePUMP Manual
MICRO-ePUMP Quick Start Guide
MICRO-ePUMP Brochure
- ePUMP Capillary Kit300744For pricing, Customers outside of the US and Canada, please contact your distributor.
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没有序列识别的功能,他就是简单的催化相邻DNA链的5-P末端和3'-OH末端以磷酸二酯键结合的反应,需ATP作辅酶。 不仅可以催化粘性末端之间或平滑末端之间的DNA的连接,也催化DNA与RNA之间以及少数RNA之间的连接。
反转录酶(Reversetranscripatase)是以RNA为模板指导三磷酸脱氧核苷酸合成互补DNA(cDNA)的酶。哺乳类C型病毒的反转录酶和鼠类B型病毒的反转录酶都是一条多肽链。鸟类RNA病毒的反转录酶则由两上亚基结构。真核生物中也都分离出具有不同结构的反转录酶。
在进行RT反应之前,应考虑以下几个方面:
1、RNA
成功的cDNA合成来自高质量的RNA,高质量的RNA至少应保证全长并且不含逆转录酶的抑制剂,如EDTA或SDS。在提取RNA的过程中,要特别防止RNase的污染,同时在逆转录反应中经常加入RNase抑制剂以增加cDNA合成的长度和产量。RNase抑制剂要在第一链cDNA合成反应中,在缓冲液和还原剂(如DTT)存在的条件下加入,因为cDNA合成前的过程会使抑制剂变性,从而释放结合的可以降解RNA的RNase。蛋白RNase抑制剂仅防止RNaseA,B,C对RNA的降解,并不能防止皮肤上的RNase,因此尽管使用了这些抑制剂,也要小心不要从手指上引入RNase,实验过程中经常更换新手套。
2、引物的选择
OligodT
选择OligodT时,要求RNA必须有PolyA,所以真核生物的mRNA都适用。适合长链甚至全长mRNA的RT,所以对RNA样品的质量要求较高,最好不要有明显的DNA污染、RNA降解和RNA断裂。假如想探索新的mRNA进行RT反应,建议推荐使用OligodT引物。使用OligodT引物要比随机引物和特异性引物的稳定性要好。
随机引物
适合各种RNA的RT,尤其适合模板丰度很低的情况(比如某个gene表达量很低)。选择随机引物时,第一链cDNA合成反应中就是以所有的RNA为模板,然后进行PCR反应时设计引物进行特异性扩增。同时要注意随机引物的量和总RNA量之间的关系,一般建议每5μg总RNA的随机引物的用量为50ng,如果每5μg总RNA的随机引物的用量超过250ng,可能会导致小片段产物(<500bp)的增加和长片断、全长产物产物的降低。
特异性引物
特异性引物只能用你设计引物时的下游引物做RT,引物设计质量影响RT的结果,而且不同引物退火温度本来就不相同,所以按照说明书按照一个温度做不是最佳选择,一般不推荐。向左转|向右转
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