Search
Search
查看更多产品信息 BenchPro™ 2100 Piercing Device - FAQs (MC3001)
67 个常见问题解答
在运行中出现这种情况是完全正常的，我们经常看到，但是它不会对仪器运行带来任何问题。你可以使用穿刺工具将试剂槽顶部的孔扩大，这有助于让液体回流到试剂槽内。如果向系统中加入了过多OD值很高的培养物，则试剂槽铝膜的顶部会出现更多的固体残留，因此，检查是否是这种情况并减少菌液使用量。在长时间使用仪器之后，你可能会看到在抽屉的顶部甚至有时在在仪器的轨道上出现固体残留物，但是这不会损伤仪器。但是，如果这种情况确实造成了困扰，我们建议使用一些Kimwipes纸巾，将纸巾用70%乙醇润湿，然后将残留物擦拭除去。
请按顺序进行以下步骤以解锁抽屉：
1.关闭仪器电源。
2.移除纯化卡（如果已经插入的话）。
3.向内按压抽屉（按住仪器的后部）以确保锁定装置已经松开，然后放松压力。
4.打开仪器电源。
5.再次向内按压抽屉（按住仪器的后部）试图将门打开。
软件在重启过程中会向门锁定电磁阀发出解锁信号。如果在重复步骤1到5数次后门仍然不能打开，请联系我们的技术支持团队。
这通常是由于空气流速的突然下降（空气系统的问题，通常不是仪器自身的问题）引起的。在BenchPro 2100运行程序中，裂解步骤和洗脱步骤需要较高的空气气压，所以这一错误信息通常出现在这两个步骤。如果这一错误出现在裂解步骤，我们建议从头重新开始，使用新的菌液，新的纯化卡，以及新的试剂槽。原样本将损失。
如果这一错误出现在洗脱步骤，可以看看是否得到了一些洗脱下来的DNA。希望有一些DNA已经被洗脱下来（通常大部分DNA在洗脱步骤的前1-2分钟已经洗脱下来）。如果这一错误正好在洗脱步骤开始前出现，而洗脱管内还没有任何溶液，你可以清空所有试剂槽并重新在正确的试剂槽内加入1.5 ml TE或水，然后使用同一试剂槽和纯化卡设置程序并重新和运行实验步骤。
这些错误信息是阀门错误信息。错误编码表明哪个阀门坏了。
通常，"valve=1/Error Code 52=1”意味着空气供应问题，因为阀门1（valve1）通常会因为空气气压或流速过低而出现故障。如果你看到这一错误信息，请测量空气气压和流速。一旦确认，关闭电源并重启机器。仪器将通过自检测试。但是，如果当空气气压和流速满足仪器要求时问题仍然存在，则意味着仪器需要修理，因为空气管线内存在液体或固体残留。造成此问题的最常见原因是使用了过高营养的培液或过多的细菌量。 对于 “valve=5/Error Code 52=5”或其它任何编码不是1的错误信息，可以尝试重启仪器。如果同样的阀门错误再次出现，通常意味着这个特定的阀门出现故障。如果在重启之后，你看到一个不同的错误编码，请检查空气气压和流速是否满足仪器要求。出现这一现象可能有以下三个原因：
空气流动的变化会降低从沉淀膜上回收的质粒DNA量。预期的回收洗脱体积应该在800 至1000 µl之间。
如果纯化的是很大的质粒，那么DNA的大小可能会阻碍从沉淀膜上回收洗脱液。我们已经确认，如果质粒很大或其大小超出了推荐的范围，那么洗脱体积（和产量）将会减小。
如果你使用丰富培养基（TB，Circlegrow等），那么洗脱体积过低可能是由于系统过载从而导致在前几个循环中真空泵的负载过高，试剂吹回到试剂托盘，吹出一些洗脱体积。房间气压降低也会导致洗脱体积减少。
产量是可能变化的，但是每次纯化应当在500 µg至1 µg之间。这一范围是由进入仪器的空气气压和气流，细菌量（由于体积、菌种、培养基类型不同而不同），菌液生长时期，质粒大小，以及质粒上的ori（复制起始位点）导致的。
最常见的两个导致低产量/零产量的原因是：低空气气压和/或低流速（仪器要求空气流速在90 psi时达到4 cfm）以及菌液过量（我们建议使用100–125 mL OD 2.0的LB培养物）。
不能，目前版本的软件不允许打断程序并继续运行。
不能，目前版本的 BenchPro 2100不能和任何实验室信息管理系统进行联接。
是的。你可以使用LB培养液或灭菌水重悬该细胞沉淀，使得其600 nm OD值为2.0。
不能。因为BAC克隆太大，所以该仪器不适合提纯BAC克隆。BenchPro2100仪器能够纯化2.7至20 kb大小的质粒。
我们已经测试了DH10B-T1R, DH5α, JM109, TOP10, 及XL1-Blue，预想所有由上述菌种衍生出来的菌种都可以使用。注意：endA+菌种是可以使用的，但是在最终纯化得到的质粒内可能含有核酸酶活性。
要获得最好结果，请参照下列指导：
•使用密度大约为109 cells/mL或600nm吸光度为2.0 至2.4的菌液。使用处于从对数生长期转向平台期的菌液。
•使用高拷贝质粒以获得高产量的质粒DNA。对于高拷贝质粒，每毫升过夜培养的LB菌液通常可以得到2–6 µg DNA。
你可以使用湿布擦拭BenchPro 2100仪器表面。不要使用强力的试剂或溶剂清理仪器。清理仪器抽屉的底部时可以使用70%乙醇对其喷雾，然后用纸巾擦拭去除乙醇。
该仪器纯化的质粒纯度很高，适用于所有下游应用，包括那些对纯度要求很高的应用，例如转染哺乳动物细胞，自动化或手动的DNA测序，PCR扩增，体外转录，细菌转化，克隆，以及标记。
平均而言，该仪器可以处理的生物量湿重是0.7到0.9 g。但是，根据所用细胞的不同大小和不同种属，这一指标会有差异。
是的。使用100mL的LB培养液重悬细胞团块，然后将其加入细胞内衬槽。
该仪器可以纯化2.7到20 kb大小的质粒。
BenchPro 2100仪器需要的人工设置时间小于5分钟。在设置完成后，仪器大约需要90分钟完成整个纯化过程。在纯化过程中你不需要守在旁边。
是的，你可以使用空气压缩机。请联系技术支持团队获取推荐的型号。空气压缩机需要满足下列条件：
•常规压力输出在80到100 psi之间
•在80 psi测量时，常规压力输出流速>3 cfm
•常规输出气流露点温度<5°C
BenchPro 2100质粒纯化卡和BenchPro 2100试剂槽必须一直保存在原装盒子内以防止卡嘴或槽膜遭到破坏。保存温度应为室温。
不能。细胞内衬槽不能重复使用。它们需要在每次程序结束后丢弃。每个BenchPro 2100试剂盒都提供相应的内衬槽和盖子。细胞内衬槽目前不能单独购买。
废液槽可以重复使用，并且它是由可以进行高压灭菌的材料制成的。你也可以订购新的废液槽。
不能，BenchPro 2100质粒纯化卡是不能重复使用的。在纯化过程中，E. Coli菌体被捕获在纯化卡内并裂解，所以纯化卡不能重复使用。
BenchPro 2100质粒纯化工作站可以同时处理一个或两个样本。
不能，该仪器使用正压工作，不能用负压工作。它使用室内空气管道工作，它是一条橙色的强制输出空气的管道。气压必须是80–100 psi。
你将需要一个室内的空气输出口（80–100 psi, 3 cfm, 稳定气流速率），或者一台相同规格的压缩机，用来连接到BenchPro 2100质粒纯化工作站。
你可以使用一个Hedland气压计测量气流速率。最低的标准是3立方英尺每分钟。气流速率没有上限。
仪器每次运行都要进行一次自我检测，每次大约需要6分钟。
你可以移出700µL TE，从而使最终的DNA浓度更高。我们不建议使用少于750 µL TE缓冲液洗脱。
对于LB培养基，你可以向菌液槽内装入最大体积150mL的菌液。对于TB培养基，虽然我们没有测试过，但是根据用户的反馈，将50mL菌液用50 mL培养液稀释后抽提的效果也很好。
虽然根据您的质粒的大小和拷贝数的不同以及菌种的不同，产量和纯度会有差异，但是我们在几种常用菌种包括C600, DH10B/T1R, DH5alpha, JM109, TOP10, 及XL1-Blu中测试了一种高拷贝质粒（pcDNA3.1），证明平均产量超过500 µg。
是的，我们提供BenchPro 2100质粒纯化系统，它是一个完全自动化的大抽纯化系统。
It is completely normal for this to happen during the run and we often see it, but haven't had it lead to any problems in the instrument. You can try widening the holes with the piercing tool, which will help any of the liquid run back into the tray. More debris on the top of the foil can occur when there is too much of a high OD culture added into the system; therefore, check this and scale back accordingly. After heavy use of the instrument, you may see some of this debris ending up in the drawer and occasionally on the tracks of the instrument, but it won't hurt the instrument. However, if it's a concern, we would recommend taking some Kimwipes wipers, moistening them with 70% ethanol, and wiping it down.
Please follow the sequence of steps below to try to release the drawer lock:
- Power-off the instrument.
- Remove card (if already inserted).
- Press the drawer inward (holding the back of unit) to ensure the lock mechanism is not catching, then release pressure.
- Power-on the instrument.
- Press the drawer inward (holding the back of unit) again and attempt to open door.
- The software sends an unlock signal to the door lock solenoid during the start-up process. If after a few attempts of steps 1 through 5 the door still does not open, please call our Technical Support team.
This is usually caused by a sudden dip in the air flow (an air system problem, usually not related to the instrument itself). During the BenchPro 2100 procedure, the lysis step and elution step are the two steps that require higher air pressure, so this error message is usually seen at these two steps. If this error happens during the lysis step, we recommend restarting from the beginning, with a new culture, new card, and new reagent tray. There is no way to recover the sample.
If this happens at the elution step, you can see if you can get any eluted DNA. Hopefully some DNA has already eluted out (usually most DNA is eluted out in the first 1-2 minutes of the elution step). If this happens right before the elution step, with no solution in the elution tube yet, you can empty out the reagent tray reservoirs and refill them with 1.5 ml of TE or water in the right reservoir, then set up the run with the same tray and card and run the protocol again.
These are valve errors. The error number tells which valve is failing.
Typically, “valve=1/Error Code 52=1” means an air supply issue, as valve 1 often fails if the air pressure or flow is on the low side. If you see this error message, please measure the air pressure and flow rate. Once verified, cycle the power and restart the machine. It should pass the self-check test. However, if the problem persists even if the air pressure and flow rate meet the instrument's needs, it means the instrument needs to be serviced due to liquid or debris in the air lines. The most common cause for this failure is using too-rich medium or too many cells.
For “valve=5/Error Code 52=5” or any number except 1, try to reboot the instrument. If the same valve error shows up, it usually suggests that this specific valve is failing. If after rebooting, you see a different error number, please check to see if the air pressure and flow rate meet the requirements.
There are three possible reasons for this:
- The variability of the air flow will reduce the amount of plasmid DNA recovered from the precipitator membrane. An expected elution volume recovery should be between 800 and 1,000 µl.
- If larger plasmids are in use, the DNA size may impede the precipitator membrane enough to reduce the final elution flow. We have verified that elution volume (and yield) may be reduced if plasmids are large or exceed the recommended range.
- If you using enriched medium (TB, Circlegrow, etc.), the low elution volume may be due to overloading of the system, causing the pump to overload during the first few pumping cycles, causing blow-back into the reagent tray and blowing out some of the elution volume. A drop in house air pressure can also cause a low elution volume.
Yield can be variable, but should be between 500 µg and 1 µg per purification. This range is a result of: air pressure and air flow into the instrument, cell amounts (due to volume, strain, or type of medium), culture growth phase, plasmid size, and the ori on the plasmid.
The two most common seen causes for low/no yield are: low air pressure and/or flow rate (the instrument requires air flow rate of 4 cfm at 90 psi) and overloading (we recommend using 100-125 mL of LB culture at OD 2.0).
No, the current version of your software will not allow you to interrupt and subsequently continue your run.
No, the current version of the BenchPro 2100 cannot be connected to any laboratory information management systems.
Yes. You can resuspend the pellet in LB media or sterile water such that the OD at 600 nm is 2.0.
No. Purification of BAC clones is not optimal due to their large size. The BenchPro 2100 instrument is capable of purifying plasmids from 2.7 to 20 kb.
We have tested DH10B-T1R, DH5alpha, JM109, TOP10, and XL1-Blue in-house, and expect that any strains derived from these will also work.
Note: endA+ strains have been shown to work, but there may be nuclease activity in the final purified plasmid.
For best results, follow these guidelines:
- Use a bacterial culture at a cell density of approximately 10E9 cells/mL, or an optical density of 2.0 to 2.4 at 600 nm (OD600). Use bacterial culture in transition between exponential and stationary phase.
- Use a high copy number plasmid to obtain a good yield of plasmid DNA. High copy number plasmids typically yield 2-6 µg DNA/mL LB culture grown overnight.
You can clean the surface of the BenchPro 2100 instrument with a damp cloth. Do not use harsh reagents or solvents to clean the unit. Clean the bottom of the instrument drawer with a light spray of 70% ethanol and then wipe off the ethanol with a paper towel.
The purified plasmid is ultrapure and suitable for all downstream applications, including those requiring the highest purity, such as transfection of mammalian cells, automated and manual DNA sequencing, PCR amplification, in vitro transcription, bacterial cell transformation, cloning, and labeling.
On average, you can process 0.7 to 0.9 g of wet pellet biomass on this instrument. However, this is subject to variability when working with different strains.
Yes. Resuspend the cell pellets in 100 mL of LB medium before you load the mixture onto the cell liner tray.
The instrument is capable of purifying plasmids from 2.7 to 20 kb.
The BenchPro 2100 instrument needs a manual setup time of less than 5 minutes. After setup, the instrument takes about 90 minutes for the entire purification process. You do not need to be present during the purification process.
We offer an Air Compressor (Cat. No. MC5001), which is validated for use with the BenchPro 2100 Plasmid Processing Station.
Air compressors from other vendors could be used too, if they meet the following requirements:
- Regulate pressure output between 90 and 120 psi
- Regulate pressure output flow at greater than 4 cfm, when measured at 90 psi
- Output clean and dry air constantly
Always store the BenchPro 2100 Plasmid Card and BenchPro 2100 Reagent Tray in the supplied box to prevent any damage to the card sippers or to the tray foil. Store both at room temperature
No. The cell liner tray and lid are not reusable. They should be disposed of after each procedure. Each BenchPro 2100 reagent kit comes with a supply of cell liner trays and lids. Cell liner trays are not currently available for standalone purchase.
The waste tray is reusable and is made of material that can be autoclaved. You can also order new waste trays.
No, the BenchPro 2100 Plasmid Purification Card is not reusable. During the purification, E. coli cells are captured and lysed within the card, making subsequent reuse impossible.
You can run one or two samples simultaneously on the BenchPro 2100 Plasmid Processing Station
No, the instrument works using positive air pressure, and cannot work on vacuum. It works using house air, which is the orange line in the lab that forces air out.
You will need an in-house air supply outlet (90-120 psi, > 4 cfm, constant flow rate), or a compressor unit with the same specifications, that can be connected to the BenchPro 2100 Plasmid Processing Station.
You can use a Hedland air gauge to measure the air flow rate for the BenchPro 2100 Plasmid Purification instrument.
A self test is performed each time a run is started and takes approximately 6 minutes to complete.
Yes, you can replace TE with water. Carefully remove all TE solution from the TE-containing well (the one closest to the elution tube slot on the reagent tray), rinse the well twice with purified water and then fill with 1.5 mL purified water. You can also pipette out up to 700 µL of TE, so that the final DNA concentration could be higher. We would not recommend using less than 750 µL TE buffer.
We recommend to use 100-125 mL of LB culture at about OD 2.0. We don't recommend using enriched media like TB, YT, etc., as a high cell density will clog the membrane and decrease DNA yield from the purification. If you already have culture in TB, dilute 50 mL culture wtih 50 mL media.
While yield and purity will differ depending on the size of your vector and copy number as well as cell strain used, a high copy vector (pcDNA3.1) has been tested in several popular strains including C600, DH10B/T1R, DH5alpha, JM109, TOP10, and XL1-Blu, demonstrating an average yield above 500 µg. Purity-wise, isolated DNA usually have both A260/A280 and A260/A230 ratios in the range of 1.8-2.0. Endotoxin is usually < 10 EU/µg (3 EU/µg average).
Yes, we offer the BenchPro 2100 Plasmid Purification System, which is a fully automated maxiprep purification system.
We recommend only running one BenchPro 2100 Maxi Plasmid Processing Station instrument for each air compressor.
The minimum specification for the BenchPro 2100 Maxi Plasmid Processing Station air flow rate is 3 CFM. There is no maximum air flow rate specified.
There are 5.58mL of isopropanol and 5.33mL of 70% ethanol per tray in the BenchPro 2100 Plasmid Purification Card and Reagent Tray Kit.