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查看更多产品信息 KingFisher™ Flex Purification System, KingFisher with 96 Deep-well Head - FAQs (5400630)
109 个常见问题解答
您可以在产品页面找到MagMAX mirVana总RNA提取试剂盒在MagMAX Express-96, KingFisher Flex, 或KingFisher Duo仪器上使用的脚本。
不是，处理体积指的是24孔深孔板内可以装入的最大体积。这包括结合缓冲液的体积和磁珠体积，所以当扩大反应体积时，一定要确保总体积不超过5mL。
不能，一旦实验程序停止，它将从头重新开始运行。您可以使用磁力架手动完成剩下的提取程序，或者从实验停止的步骤开始重新创建一个修改版的KingFisher Flex实验程序并运行该程序。
在BindIt软件中设置实验方案。在完成前8块板后，仪器将暂停并提示您将板换出。
转移锁仅在移动仪器时有必要使用。
我们强烈建议保持每孔的液体体积在指定的限值范围内，以免试剂溢出并确保最佳结果。
这种情况有时会发生，但是这不会影响产量，因为样本已经从磁珠上被洗脱下来了。
如果起始材料过于粘稠，那么磁力棒将不能完全吸取磁珠。可以尝试稀释样本，并检查样本是否使用正确的方法进行匀浆及裂解；向样本内加入少量去垢剂也可以提高磁珠的回收率；还可以对样品板进行快速离心，使磁珠沉积在板的底部，以提高磁珠的回收率。
如果没有向磁棒套架插入磁棒套，则程序不会开始运行。
将软布或纸巾用温和的去垢剂、肥皂水或酒精润湿，然后擦拭磁力棒。
加热模块每分钟升温10°C，因此需要6分钟左右。
您可在同一次运行中使用深孔板和KingFisher 96孔板。因此，您可以先采用较大体积的反应板开始处理，之后将纯化样本洗脱到小体积反应板（KingFisher 96孔板或MagMAX Express-96孔板）中。
可以通过运行“维护 (Maintenance)”菜单之下的“更换磁头 (Change magnet)”程序实现，使用“上 (up)”和“下 (down)”按钮进行选择。
The KingFisher Flex磁体是使用非常稳定的材料制成的。磁场不会变弱。不过，极端的机械外力或加热可能会对磁体造成损坏。
注：务必使KingFisher Flex的磁棒头远离其他磁棒或其他磁体。磁体相互接触会引起严重损坏。KingFisher Flex使用一个USB接口，而MagMAX Express- 96需要一个RS-232接口。
实验方案不能从KingFisher Flex仪器转移到电脑。实验方案需要被存储在BindIt软件内之后才可以进行编辑。
是的，MagMAX Express-96仪器可以和装有最新版本BindIt软件v3.3.1.7的电脑相连。这种连接需要一个到USB转化器的串行端口。已有的.kf2 文件在BindIt软件内不能被编辑，但是可以将.kf2文件重写为.bdz文件从而可以被编辑。请参见应用注意（https://tools.thermofisher.com/content/sfs/brochures/KingFisher-BindIT-TechNote-5189009.pdf）了解更多细节。
不能，出厂实验方案不能编辑，但是您可以使用“另存为”功能对其重命名，然后将它作为一个新的版本来编辑。
BindIt软件和Windows XP (Professional with Service Pack 3)，Windows Vista (Service Pack 2), Windows 7 (Service Pack 1), 以及Windows 8兼容。
对于核酸提取，我们建议使用我们的MagMAX试剂盒和MagJET试剂盒。其它大小在0.5到10 µm间的磁珠应该也可以使用。但是，您需要自己用Thermo Scientific BindIt软件编写您自己的KingFisher实验方案，并自己验证/优化这一实验方案。
可以，可将样品分装。在进行下一步之前，设置程序使得磁头可以从多个孔内收集样本。确保不要过量加样。
可以，反应体积可以扩大，并且所有其它体积（例如洗涤体积）需要以相同倍数扩大。除非板的容积允许，我们强烈建议反应体积扩大不应超过2倍。
不行，我们强烈推荐您使用MagMAX板或KingFisher板，因为它们是专为配合KingFisher Flex磁棒套同时使用而设计的，以达到最佳效果。其它制造商生产的板子可能与KingFisher Flex加热模块不兼容。它们还可能引起意料之外的问题，例如可能会由于不同的孔容积和板底高度而引起交叉污染。
带96深孔磁棒头的KingFisher Flex仪器和MagMAX Express-96仪器所需的耗材是相同的。列表如下：
- MagMAX Express-96 深孔板 (货号 4388476)
- MagMAX Express-96 标准板(货号4388475)
- MagMAX Express-96深孔磁棒套(货号4388487)
- MicroAmp 透明封口膜（可选）： (货号. 4306311)
对于带24深孔磁棒头的KingFisher Flex仪器，所需耗材列表如下：
- KingFisher Flex 24深孔板：(货号. 95040480)
- KingFisher Flex 24深孔磁棒套： (货号 97002610)
不可以，96孔标准磁棒头和96深孔磁棒头不能互换。但是，两台仪器可以使用相同的磁棒套。
KingFisher Flex 磁珠处理仪有四种磁棒选择：
- 用于MagMAX Express-96深孔板的KingFisher Flex磁棒
- 用于KingFisher 24深孔板的KingFisher Flex磁棒
- 用于KingFisher 96孔板的KingFisher Flex磁棒
- 用于有裙边PCR板的KingFisher Flex磁棒
是的，所有MagMAX试剂盒都能用于KingFisher Flex 磁珠处理仪。
Once you place your order, please contact our Instrument Service team at 1-800-955-6288 option 3, 1 (this kit is only available in North America). You will be given an access code to the TaqPath Monkeypox/Orthopox Virus DNA Kit eLearning where you’ll learn how to install, update, and download the Pathogen Interpretive Software package for the TaqPath Monkeypox/Orthopox Virus DNA Kit.
Results are analyzed using the following software:
- Applied Biosystems Pathogen Interpretive Software v1.1
- SAE Administrator Console Dx v1.0 or v1.2 (for security and audit functions)
- Pathogen Interpretive Software 1.0.0 DAT (SAE Profile)
Please use the appropriate assay panel and template for your instrument:
- MPXOPX-EUA_QS5-9601_1.0.0.zip
- MPXOPX-EUA_QS5Dx-9602_1.0.0.zip
- MPXOPX-EUA_QS7F-384_1.0.0.zip
Note: These zip files are located in EducationConnect (www.thermofisher.com/educationconnect). The eLearning course needs to be completed to be granted access.
To obtain the correct analysis software or firmware version for your real-time PCR instrument, go to thermofisher.com/validatedfirmware, then select your instrument in the Real-Time PCR section.
Here are the instruments that can be used with the TaqPath Monkeypox/Orthopox Virus DNA Kit:
- Applied Biosystems QuantStudio 5 Real-Time PCR System (96-well, 0.1 mL block)
- Applied Biosystems QuantStudio 5 Dx Real-Time PCR System (96-well, 0.2 mL block)
- Applied Biosystems QuantStudio 7 Flex Real-Time PCR System (384-well block)
Performance was established using the MagMAX Viral/Pathogen II Nucleic Acid Isolation Kit (Cat. No. A48383) in combination with the KingFisher Flex Magnetic Particle Processor with 96 Deep-Well Head and the KingFisher Flex 96 Deep-Well Heating Block (MVP_2Wash_400_Flex program). Other purification methods may lead to deviations in the expected result.
The TaqPath Monkeypox/Orthopox Virus DNA Kit protocol is currently authorized for use with lesion swab specimens in universal transport media (UTM) or viral transport media (VTM) from individuals suspected of monkeypox infection by their healthcare provider.
Go to www.thermofisher.com/monkeypox and download the Instructions for Use (IFU) to see the workflow outlining the instruments, consumables, and reagents required to run Monkeypox detection experiments.
The reporter dyes used are as follows:
- Monkeypox virus: FAM dye
- Orthopoxvirus: VIC dye
- RNase P: JUN dye
The TaqPath Monkeypox/Orthopox Virus DNA Kit probes contain QSY and MGB quenchers, which do not fluoresce. Select None for Quencher in the instrument set up procedure.
The TaqPath Monkeypox/Orthopox Virus DNA Kit includes the following components:
- TaqPath Monkeypox/Orthopox Virus Multiplex Assay: Multiplexed real-time PCR assay containing primers/probes specific to monkeypox virus, non-variola orthopoxvirus targets, and human RNase P.
- TaqPath Monkeypox/Orthopox Virus DNA Positive Control: Positive DNA control that contains templates specific to monkeypox virus, non-variola orthopoxvirus targets, and human RNase P regions targeted by the assay.
Note:
- The master mix is not included in the kit and needs to be purchased separately: TaqPath BactoPure Microbial Detection Master Mix (No ROX) (Cat. No. A52703, A52704, or A52705)
- TE Buffer (Cat. No. 12090015) and Nuclease-free water (MLS) are required and not supplied with the kit.
The User Manual for the BindIt Software can be downloaded from the BindIt Software product page online, under the Documents & Downloads section. It is also included within the BindIt Software installation file under "Documentation".
Find additional tips, troubleshooting help, and resources within our Automated Nucleic Acid Purification Support Center.
To determine the cause of the connection issue, we recommend checking the following:
1. Check software settings:
• Make sure the instrument selected is KingFisher Flex.
• Make sure the instrument serial number was input with the dash included and no spaces (ex, "711-xxxx").
• Make sure the communication tab was set to "USB" and not "COM".
2. Check the instrument and computer:
• The KingFisher Flex instrument screen (GUI) should be set to the "Factory Protocols" tab (blue tab) for best results.
• Make sure the port function on the Maintenance Tab (last tab) is checked for "USB".
• Try using different USB cables.
• Try a different USB port on the computer.
• Check the Windows version you are using.
• Reboot the KingFisher Flex instrument and the computer.
• Make sure the driver was installed properly. This may require the help of your local IT team.
If the connection issue persists, please contact our Technical Support team at techsupport@thermofisher.com.
Find additional tips, troubleshooting help, and resources within our Automated Nucleic Acid Purification Support Center.
You can use any brand of UPS as long as it is compatible with the following KingFisher Flex instrument electrical specifications:
• Mains power supply: 100–240 Vac, 50/60 Hz, nominal automatic voltage detection
• Power consumption: 175 VA max.; 55 VA standby
Find additional tips, troubleshooting help, and resources within our Automated Nucleic Acid Purification Support Center.
The “error message 12” error usually means there is something out of place on the KingFisher Flex instrument. We recommend the following to resolve the issue:
1. Check the heat block and make sure that it is not lifted up out of the base of the unit. If it is out of the base, check to see if there is anything underneath it that is preventing it from going down. If there is nothing under the heat block, it may have gotten stuck in the up position. This will require a service call.
2. Check that the tip shield is off to the right and out of the way from underneath the magnetic head. If it is stuck in the incorrect position, this may require a service call.
3. If the turntable was moved and it clicked out of place, the rectangular footprint cannot be centered below the head. Turn off the instrument and turn it back on. The instrument should re-align itself.
If the error continues to display, please contact techsupport@thermofisher.com for assistance.
Find additional tips, troubleshooting help, and resources within our Automated Nucleic Acid Purification Support Center.
Please follow these steps to load a script from your computer to the KingFisher Flex or KingFisher Duo/Duo Prime instrument:
1. Use the USB cable to connect the PC to the instrument.
2. Turn on the instrument.
3. Open the BindIt software and click the Connect button.
4. Select "Find New Instrument." BindIt will automatically connect to the instrument and the instrument screen will say "Computer Control".
5. Click the "Transfer" Button.
6. Click the "Upload" button.
7. Browse to find the file and double-click it.
8. In the new window that opens:
a. KingFisher Flex: you will be prompted to enter a new name – this is not necessary if the name only contains letters, digits, dash or underscore.
• If the name contains spaces, apostrophes or any other character, you should enter a new name containing only letters, digits, dash (-) or underscore (_).
• Click the "Category" pulldown to select the instrument folder you want to load the script to.
• Click OK.
b. KingFisher Duo/Duo Prime: The KingFisher Duo and Duo Prime instruments accept all characters in the filename.
• Click the "Category" pulldown to select the instrument folder you want to load the script to.
• Click OK.
Find additional tips, troubleshooting help, and resources within our Automated Nucleic Acid Purification Support Center.
We recommend that people with pacemakers consult with their doctor before working with the KingFisher instruments. For the 24 Deep-Well Head, the magnetic force is 3,500 gauss. For the 96 Deep-Well Head, the magnetic force is 1,700 gauss. For the other magnetic heads, the magnetic force is lower.
Find additional tips, troubleshooting help, and resources within our Automated Nucleic Acid Purification Support Center.
The KingFisher Flex and KingFisher Duo instruments no longer come with pre-installed scripts. If your instrument begins with the following serial numbers, it will not have pre-loaded scripts:
• KingFisher Flex serial number 711-1951 or greater
• KingFisher Duo Prime serial number 706-1366 or greater
Find additional tips, troubleshooting help, and resources within our Automated Nucleic Acid Purification Support Center.
The accuracy of the KingFisher Flex heating block is ± 1°C for up to 80°C, and ± 2°C for up to 115°C (see page 72 of the manual). If there is an issue with the accuracy of the heating block, you can contact to schedule a service call. Calibration of the heating block can only be performed by one of our engineers.
Find additional tips, troubleshooting help, and resources within our Automated Nucleic Acid Purification Support Center.
No, scripts cannot be transferred from the KingFisher Flex instrument to the computer for editing. It is possible to transfer a backup copy of the script from the KingFisher Flex instrument to the computer, but you will not be able to edit it. You may use this backup file to write a new script. In general, we recommend storing protocols in the BindIt software on the computer so the protocols can be edited there.
Find additional tips, troubleshooting help, and resources within our Automated Nucleic Acid Purification Support Center.
A list of necessary plastic consumables for the KingFisher Flex 96 Deep-Well Head can be found on the KingFisher Plastics for 96 deep-well format product page.
No training is provided with the purchase of a KingFisher Flex or KingFisher Duo Prime instrument, but you can purchase training separately. We offer onsite half-day and full-day trainings for the KingFisher Flex and KingFisher Duo Prime instruments, depending on your needs. Please contact your local instrument sales representative to obtain a quote. The training will be led by your regional Field Application Scientist.
Find additional tips, troubleshooting help, and resources within our Automated Nucleic Acid Purification Support Center.
We cannot write a script for a purification kit for another company, but we can provide you with a tutorial on how to use the BindIt software. We also recommend checking with the manufacturer of the purification kit you are using, since some companies may already have scripts for their products to be used on multiple instruments.
Find additional tips, troubleshooting help, and resources within our Automated Nucleic Acid Purification Support Center.
Scripts for processing large volumes of whole blood (up to 1 mL) can be found on the MagMAX Multi-Sample Ultra kit product page under the Product Literature section at the bottom of the page. There are two scripts available:
• KingFisher DUO file: A25597_Large_Volume_Blood.bdz
• KingFisher Flex file: A25597_Large_Volume_Blood.bdz
*Use the page scrolling function to see the complete list of product literature for this product.
Find additional tips, troubleshooting help, and resources within our Automated Nucleic Acid Purification Support Center.
We offer onsite half-day and full-day trainings for the KingFisher Flex instruments, depending on your needs.The training will be led by your regional Field Application Scientist. You can contact your local sales representative to obtain a quote for the training cost.
Find additional tips, troubleshooting help, and resources within our Automated Nucleic Acid Purification Support Center.
If the tip does not load correctly onto a Thermo Scientific KingFisher magnetic head, we recommend that you try manually flexing the tips both lengthwise and widthwise to level the tip comb. A video of flexing the tip comb can be found here: https://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/automated-purification-extraction/kingfisher-systems/resources.html. The video is under "KingFisher Flex" and titled "How to install the magnet head and check the tip comb." Flexing of the tip comb is demonstrated at timepoint 1:35. If that does not work, try a different box or lot of tips. This applies to all KingFisher instruments.
We recommend using the MagJet Plasmid DNA kit (Cat. No. K2791) for plasmid purification. Scripts for using the MagJet Plasmid DNA kit with the KingFisher Flex and KingFisher Duo Prime Purification Systems are available for download on the kit product page (https://www.thermofisher.com/order/catalog/product/K2791). The ChargeSwitch Plasmid ER Mini Kit (Cat. No. CS10100) is also an option for plasmid purification. There is no script currently available for using ChargeSwitch Plasmid ER Mini Kit, but you can create a custom protocol for use with the KingFisher Flex and KingFisher Duo Prime Purification Systems.
We do not recommend placing the KingFisher Flex Purification System in close proximity to magnetic tapes, computer discs or other magnetic storage systems, such as credit cards, as these can be damaged by the strong magnetic field of the KingFisher Flex magnetic heads. Do not hold the KingFisher Flex magnetic heads close to a PC display, since this may cause damage to the display. Do not use metal tools when handling KingFisher Flex magnetic heads. People wearing a pacemaker or metallic prostheses should also not use this product as these items could be affected or damaged by close contact with a strong magnetic field.
We recommend cleaning the KingFisher Flex instrument cover with water containing a mild soap or a slightly acidic, neutral or slightly alkaline detergent. You can use up to a 3.5% Sodium Hypochlorite concentration. We do not recommend using phenol or chloroform to clean the cover.
The KingFisher Flex Purification System cover is made of polyethylene terephthalate (PET).
The KingFisher Flex Purification System heating block is located inside the instrument and can be used automatically during the protocol. All KingFisher Flex plates can be heated using specially designed, interchangeable heating blocks. Any number of heating steps can be added to a protocol. When a protocol enters a heating step, the plate is automatically moved to the dedicated heating position for heating. Afterwards, the protocol continues automatically to the next step.
The KingFisher Flex instrument with the 24 Deep-Well magnetic head can process larger volumes for each sample (up to 5mL total volume) compared to the KingFisher Flex instrument with the 96 Deep-Well magnetic head.
Yes, the KingFisher Flex instrument 96-well magnetic head and the MagMAX Express-96 instrument magnetic head can use the same tip combs and KingFisher plates even though the magnetic heads are different.
The MagMAX Express-96 protocol scripts can be found on the MagMAX Express protocol page (https://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/rna-extraction/rna-types/total-rna-extraction/magmax-technology/magmax-particle-processors/magmax-instrument-protocols-scripts.html). If the specific script you are looking for is not located here or on the product page for the MagMAX kit, please contact Technical Support at techsupport@thermofisher.com.
We recommend using the BindIt Software to transfer scripts to the MagMAX Express-96 and KingFisher Flex instruments. The BindIt Software should be used to transfer scripts for all of the Applied Biosystems MagMAX Express and Thermo Scientific KingFisher instruments.
The only way is to do this is by rewriting the script from where you want it to start, or start the run all over again.
There is no way to edit a BindiT protocol that has been installed on the KingFisher Flex/KingFisher DuoPrime instrument. The only way to edit the protocol is by saving the original .bdz file to a PC and then editing it in BindiT software.
The KingFisher Flex instrument needs to be connected to the computer/laptop and recognized by it. The lower bottom screen should say which instrument it is connected to. If it says anything about “simulator”, it is not connected to a real instrument.
You need to add a new instrument to the database in order to recognize the instrument: Settings > Instrument Setup > New > select what kind of instrument it is from the drop-down menu (i.e., KingFisher 96/MagMAX Express 96), give the instrument a name, and enter the serial number (make sure to keep the “-” in the middle). Once the instrument is connected and recognized, you should be able to transfer files over.
Turn the instrument on and off to see whether the error message disappears. Alternatively, do a dry run without any samples. If the issue could be consumable dependent try a new set/lot of tip combs. If the error message repeatedly occurs, please contact Technical Support (techsupport@thermofisher.com).
A .kf2 file cannot be edited in the BindIt software, but it is possible to rewrite a .kf2 file as a .bdz file so that it can be edited. Please see this application note for instructions on how to convert a .kf2 file into a .bdz file (http://www.thermoscientific.com/content/dam/tfs/LPG/LCD/LCD Documents/Application & Technical Notes/Liquid Handling and Dispensing/KingFisher-BindIT-TechNote-5189009.pdf). The speed settings are different in the two software (KingFisher software has 13 speed settings and BindIt software has 5 speed settings), so the application note has a conversion table for what speed to use in the BindIt software.
They are created by different software (BindIt and KingFisher software, respectively).
.kf2 files can only be used for the MagMAX Express instrument whereas .bdz files can be used for MagMAX Express and KingFisher Flex/Duo instruments (but scripts need to be specifically written for a particular instrument).
The old KingFisher 2.6 software only works on Windows XP, whereas the BindIt software works on Windows XP, Windows 7, and Windows 8. The speed selections are also different. The old KingFisher 2.6 software featured 13 different speed setting whereas the BindIt software features 5 mixing speeds (slow, medium, fast, bottom, and half mix). The BindIt software can run KingFisher Software 2.6 files (.kf2) just as they are, but it cannot edit the .kf2 file. Please review our Technical Note for instructions on how to convert a .kf2 file to a format that can be edited in the BindIt software. A .kf2 protocol thas has been converted into a BindIt protocol can still achieve excellent yield and purity of the extracted target. Minor optimization may be required due to the differences in mix speeds and bead collection parameters, or to minimize protocol times.
The scripts for the MagMAX CORE Nucleic Acid Purification Kit can be found here (https://www.thermofisher.com/order/catalog/product/A32702). In order to obtain the .bdz scripts for the other animal health MagMAX kits (Cat. Nos. AM1835, AM1836, AM1840, 4462359), please email technical support at animalhealthandfoodsafety@lifetech.com.
Yes, depending on the kind of protocol and head that is used, the following consumables will have to be purchased:
- KingFisher 96 deep-well plate (Cat. No. 95040450)
- KingFisher 96 deep-well plate, sterile (Cat. No. 95040460)
- KingFisher 96 tip comb for deep-well (Cat. No. 97002534)
- KingFisher 96 deep-well tip comb and plate, sterile (Cat. No. 97002820)
- KingFisher 96 microplate (200 μL) (Cat. No. 97002540)
- KingFisher 96 tip comb (Cat. No. 97002524)
- MicroAmp Clear Adhesive Film (optional) (Cat. No. 4306311
- The KingFisher Flex instrument has the option of using a 24-Deep-Well head in addition to a 96-Deep-Well head.
- The KingFisher Flex instrument has a USB communications port, whereas the MagMAX Express-96 instrument only has a RS-232 port.
- The KingFisher Flex instrument uses snap-in heat blocks, while the heat blocks need to be screwed in for the MagMAX Express-96 instrument.
- Unlike the MagMAX Express-96 instrument, the KingFisher Flex instrument has a graphical user interface and expanded keypad (LCD screen).
We recommend one of the following instruments:
- KingFisher Flex Purification System (Cat. No. 5400640)
- KingFisher Duo Prime Purification System (Cat. No. 5400110)
- MagMAX Express Magnetic Particle Processor (AM-Applied Markets Customer) (Cat. No. 4400075)
PVP adsorbs polyphenols, removing them from the final eluate. DTT, a reducing agent, protects the DNA from degradation by preventing the oxidation of polyphenols. We generally recommend adding PVP for woody, lignified, or polyphenol samples (branches, twigs, needles, and wax-coated leaves). DTT can be helpful when working with seeds. Some seeds (e.g., seeds with a pesticide coat) do benefit from addition of PVP to Lysis Buffer A. If DTT supplementation to Lysis Buffer A was already attempted and has not helped, try again with PVP instead.
If higher DNA concentration is needed, the elution volume can be reduced to 50 µL when using the KingFisher Flex instrument. However, we have not tested this and the script may have to be changed accordingly.
You can find the .bdz files on the product page (https://www.thermofisher.com/order/catalog/product/A32549?ICID=search-product).
Table 1 of the manual (https://tools.thermofisher.com/content/sfs/manuals/MAN0015954_MagMAX_Plant_DNA_Kit_UG.pdf) lists all the different plants we have tested with the kit and the approximate DNA yields to expect.
Yes, both the KingFisher Flex 96 KF and 96 Deep-Well heating blocks can be used for the 96 Deep-Well Head, but they are only compatible with certain MagMAX/KingFisher plates.
- The KingFisher Flex 96 KF Heating Block is compatible with both standard plates and Deep-Well plates (Cat. Nos. 97002540, 4388475, 95040460, 4388476).
- The KingFisher Flex Deep-Well Heating Block is only compatible with Deep-Well plates (Cat. Nos. 95040460, 4388476).
Yes, here are the catalog numbers for standalone heating blocks:
- Cat. No. 24075420: KingFisher Flex 96 KF Heating Block
- Cat. No. 24075430: KingFisher Flex 96 Deep-Well Heating Block
- Cat. No. 24075440: KingFisher Flex 24 Deep-Well Heating Block
Yes, it comes with both the KingFisher Flex 96 KF Heating Block and KingFisher Flex 96 Deep-Well Heating Block.
No, the MagMAX Cell-Free DNA Isolation Kit contains the Dynabeads MyOne SILANE magnetic beads. For this reason we recommend using the DynaMag magnets (Cat. Nos. 12302D, 12321D).
The MagMAX Cell-Free DNA Isolation Kit has been designed to isolate cell-free DNA (cfDNA) from plasma, serum, and urine samples.
Levels of cell-free DNA in circulation can range from 1 ng/mL to 100 ng/mL of plasma or serum depending on the donor. For samples containing low levels of cell-free DNA, we recommend increasing the starting sample volume.
Yields from hemolytic plasma, lipemic plasma, or other compromised samples types vary greatly from donor to donor. We recommend processing these types of samples using the manual protocol.
You can find scripts for using MagMAX mirVana Total RNA Isolation Kit with the MagMAX Express-96, KingFisher Flex, or KingFisher Duo instrument, on the product page (http://www.thermofisher.com/order/catalog/product/A27828).
No, the processing volume refers to the maximum volume that can be held in the 24 Deep Well plates. This incorporates the volume of binding buffers and beads, so when scaling up reaction volumes, it is important to make sure that the total volume does not exceed 5 mL.
No, once a protocol was stopped, it will restart from the beginning. The extraction procedure can either be finished manually by using a magnetic stand, or a modified version of the original KingFisher Flex script would have to be created that starts off at the point where the run was stopped.
Set up the protocol on the BindIt software. After the first 8 plates are done, the instrument will pause and instruct you to switch out the plates.
The transport locks are only necessary when relocating the instrument.
We strongly recommend that you maintain the specified volumes within the defined limits to avoid spillover and to help ensure the best performance at the most efficient level.
This happens sometimes but it will not affect the yield because the sample has been released from the particles.
If the starting material is too viscous, the magnetic rods will not be able to collect the particles. Dilute the sample and check that the sample is properly homogenized/lysed. Adding low amounts of detergent will improve the collection of magnetic particles as well. Quickly centrifuge the plate to sediment the particles to the bottom of the plate.
The protocol will not start without the tip combs inserted into the tip comb holder.
Wipe the magnetic rods with a soft cloth or tissue paper soaked in a mild detergent solution, soap solution, or alcohol.
The heating block warms up at the rate of about 10°C per minute, so it will take about 6 minutes.
Both deep-well plates and KingFisher 96 plates can be used during the same run. Therefore, it is possible to start the processing by using larger volumes (in a deep-well plate) and elute the purified sample to a smaller volume (in a KingFisher 96 or MagMAX 96 standard plate).
This can be done by running the Change magnet protocol under the Maintenance menu using the “up” and “down” cursor keys.
The KingFisher Flex magnets are made of material that is very stable. The magnetic field will not get weaker. However, extreme mechanical force or heating can cause damage to the magnets.
Caution: It is very important to keep the KingFisher Flex heads away from each other and from other magnets at all times. Clashing together of the magnets may cause serious damage to the magnets.
KingFisher Flex uses a USB port, whereas the MagMAX Express- 96 requires a RS-232 port.
No. Protocols cannot be transferred back from the instrument to the computer. Protocols need to be previously stored in the BindIt or KingFisher software to make edits.
Yes, the MagMAX Express-96 instrument is compatible with the BindIt software. A serial port-to-USB converter is required to connect the MagMAX Express-96 instrument to the computer.
If you are using BindIt software v3.3.1.7 you can use it to run existing .kf2 files. You will not be able to edit the .kf2 files in v3.3.1.7.
If you are using BindIt software v4.0 or later, you cannot run or edit existing .kf2 files. To get around this, you can convert the .kf2 files to the BindIt (.bdz) file format and then edit the .bdz file in the BindIt software. Please see the following application note for instructions on how to convert the .kf2 files: (http://www.thermoscientific.com/content/dam/tfs/LPG/LCD/LCD Documents/Application & Technical Notes/Liquid Handling and Dispensing/KingFisher-BindIT-TechNote-5189009.pdf). If you have already converted your .kf2 files to .bdz files, or you plan to convert your .kf2 file to .bdz files, we recommend using BindIt software v4.0 or higher.
No, manufactured protocols are locked for editing, but they can be renamed using the ‘save as' function and edited as a new version.
BindIt software v3.3.1 is compatible with Windows XP (Professional with Service Pack 3), Windows Vista (Service Pack 2), Windows 7 (Service Pack 1), and Windows 8.
BindIT software v4 is compatible with Windows 7 with Service Pack 1 (32- or 64-bit), Windows 8.1 (32- or 64-bit), and Windows 10 (32- or 64-bit).
For nucleic acid extraction, we recommend using our MagMAX kits and MagJET kits. Other magnetic beads should work well as long as they are 0.5 to 10 µm in size. However, you will have to write your own KingFisher protocols using the Thermo Scientific BindIt software and validate/optimize the protocol on your own.
Find additional tips, troubleshooting help, and resources within our Automated Nucleic Acid Purification Support Center.
Yes, this can be done by sample splitting. Set the protocol so that the magnet will collect samples from multiple wells before continuing to the next step. Be sure not to overload.
Find additional tips, troubleshooting help, and resources within our Automated Nucleic Acid Purification Support Center.
Yes, reaction volumes can be scaled up, and all other volumes (i.e. wash volumes) will have to be increased by the same fold difference. It is generally not recommended to scale up reaction volumes more than 2-fold unless the plate's volume capacity will allow it.
No, we strongly recommend that you use the MagMAX plates or KingFisher plates since they were specifically designed to be used with KingFisher Flex tip combs to attain maximal performance. Plates from other manufacturers may not be compatible with the KingFisher Flex heating blocks. They may also cause unexpected problems, such as cross-contamination, due to the divergent well volume and bottom height of the plate.
The consumables for the KingFisher Flex instrument with the 96 Deep Well head are listed below:
- KingFisher 96 deep-well plate (Cat. No. 95040450)
- KingFisher 96 deep-well plate, sterile (Cat. No. 95040460)
- KingFisher 96 tip comb for deep-well (Cat. No. 97002534)
- KingFisher 96 deep-well tip comb and plate, sterile (Cat. No. 97002820)
- KingFisher 96 microplate (200 μL) (Cat. No. 97002540)
- KingFisher 96 tip comb (Cat. No. 97002524)
- MicroAmp Clear Adhesive Film (optional) (Cat. No. 4306311
For the KingFisher Flex instrument with the 24 Deep Well head, the following consumables are required:
- KingFisher 24 deep-well plate (Cat. No. 95040470)
- KingFisher 24 deep-well plate, sterile (Cat. No. 95040480)
- KingFisher 24 deep-well tip comb and plate (Cat. No. 97002610)
- KingFisher 24 deep-well tip comb and plate, sterile (Cat. No. 97002620)
No, the 96 well standard and 96 Deep Well heads are not interchangeable between the MagMAX Express-96 and KingFisher Flex instruments. However, the same tip combs can be used for both instruments.
There are 4 magnetic head options available for the KingFisher Flex Magnetic Particle Processor:
- KingFisher Flex head for MagMAX Express-96 Deep Well plate
- KingFisher Flex head for KingFisher 24 Deep Well plate
- KingFisher Flex head for KingFisher 96 plate
- KingFisher Flex head for PCR plate, skirted
Yes, all MagMAX kits can be used with the KingFisher Flex Magnetic Particle Processor.