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Invitrogen™ MagMAX™-96 for Microarrays Total RNA Isolation Kit

Catalog No. AM1839
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AM1839 96 preps
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Catalog No. AM1839 Supplier Invitrogen™ Supplier No. AM1839
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Description

Includes

Kit contents: 1 Processing Plate and Lid; 12mL Lysis/Binding Solution Concentrate; 18mL Wash Solution 1 Concentrate; 72.5mL Wash Solution 2 Concentrate; 14mL Elution Buffer; 6mL MagMAX TURBO DNase Buffer; 4 x 25mL TRI Reagent; 1.1mL RNA Binding Beads; 1.1mL Lysis/Binding Enhancer; 215μL TURBO DNase, 10U/μL. Contains enough reagents to isolate RNA from 96 samples.

The MagMAX™-96 for Microarrays Total RNA Isolation Kit is for the rapid, high-throughput purification of RNA from mammalian cells and tissue in 96-well plates.

The MagMAX™-96 for Microarrays Total RNA Isolation Kit is for the rapid, high-throughput purification of RNA from mammalian cells and tissue in 96-well plates. The kit combines an organic extraction with magnetic bead–based purification yielding extremely high-quality RNA suitable for the most stringent applications, such as microarray analysis. The kit includes sufficient reagents for 96 reactions. Advantages of the MagMAX™-96 for Microarrays Total RNA Isolation Kit:

• Improve gene expression studies with consistent results from experiment to experiment
• Combines TRI Reagent™ with a superior magnetic bead–based technology
• Streamlined RNA purification, without sacrificing RNA quality or quantity
• Requires less hands-on time than competitor kits
• Can be integrated with established robotic platforms
• Best MagMAX™ kit for difficult samples or large volumes

Benefits of MagMAX™ magnetic bead–based purification
Magnetic beads offer many benefits compared to other technologies for isolating RNA. Beads bind RNA more efficiently than glass fiber filters, resulting in higher and more consistent RNA yields. Additionally, because filters are not used, there is no risk of filter clogging due to cellular particulates in samples. Since only a small volume of magnetic beads is needed for high- efficiency binding, the bound RNA can be eluted in just 20–50 μL of nuclease-free water, concentrating RNA from large, dilute samples.

Highly stringent for the best-quality RNA
The MagMAX™-96 for Microarrays Total RNA Isolation Kit (patent pending) procedure combines the robust and reliable lysis/denaturant TRI Reagent™ with MagMAX™ magnetic bead–based RNA purification technology, resulting in streamlined RNA purification, even in a high-throughput format, without sacrificing RNA quantity or quality. TRI Reagent™ readily lyses even difficult cells and tissues while denaturing nucleases to maintain the integrity of RNA. The kit provides high yields of pure, intact RNA that can be used directly for quantitative reverse transcriptase PCR (qRT-PCR) and microarray analysis. See the accompanying data showing RNA yield and quality obtained when using the MagMAX™-96 for Microarrays Total RNA Isolation Kit. Also shown is data illustrating a high correlation between the method that Affymetrix recommends for producing RNA for use on their GeneChips™ and the method that the MagMAX™-96 for Microarrays kit employs.

Fast, flexible procedure
Two alternative protocols are supplied for the kit. The “Spin” method has fewer steps, requires no DNase treatment, and allows for larger sample amounts to be processed. The “No Spin” method requires less sample handling and can easily be integrated with a robotic liquid handler. The procedure is fast (less than 1 hr), simple, and well suited for automation. The kit can also be used to process fewer than 96 samples at one time and is compatible with either manual processing using multi-channel pipettors or processing with robotic liquid handlers. Downloadable protocols for the use of this kit with specific liquid handling systems are available at our Automation Resource.

Accessory products
A 96-well magnetic ring stand is required to use this kit. The Magnetic Stand-96 (Cat. No. AM10027) and the 96-well Magnetic Ring Stand (Cat. No. AM10050) are designed for paramagnetic bead precipitation from standard 96-well, U-bottom microplates. The Magnetic-Ring Stand pellets the magnetic beads in a circle shape, enabling easy removal of supernatant by a robot or multichannel pipettors. For additional processing plates, Matrix 96-Well U Bottom Polypropylene Plates (Thermo Scientific, Item# 4917) are recommended.

Specifications

Content And Storage

Kit Contents (storage):
• 1 Processing Plate & Lid (room temperature)
• 12 mL Lysis/Binding Solution Concentrate (room temperature)
• 18 mL Wash Solution 1 Concentrate (room temperature)
• 72.5 mL Wash Solution 2 Concentrate (room temperature)
• 14 mL Elution Buffer (room temperature)
• 6 mL MagMAX™ TURBO™ DNase Buffer (4°C or room temperature)
• 4 x 25 mL TRI Reagent™ (4°C)
• 1.1 mL RNA Binding Beads (4°C)
• 1.1 mL Lysis/Binding Enhancer (-20°C)
• 215 μL TURBO™ DNase, 10 U/μL (-20°C)

Contains enough reagents to isolate RNA from 96 samples.
Format 96-well Plate
Isolation Technology Magnetic Bead, Organic Extraction
Purification Time <1 hr.
Sample Type Cells, Mammalian Cells, Plant Samples, Tissue (Mammalian), Yeast
Final Product Type Total RNA
High-throughput Compatibility High-throughput Compatible, Automated Protocols
For Use With (Equipment) MagMAX™ Express 24 and 96, Robotic Liquid Handling Systems, MagMAX™ Express-96 Standard Magnetic Particle Processor
For Use With (Application) Microarray Analysis, Real-Time Quantitative PCR (qPCR), Reverse Transcriptase PCR (RT-PCR), Northern Blotting
No. of Reactions 96 Preps
Product Line Ambion, MagMAX
Quantity 96 preps
Starting Material Amount Up to 5 x10^6 cells, Up to 100 mg tissue
Target Total RNA
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Frequently Asked Questions (FAQs)

I am working with your MagMAX Microarray Total RNA Isolation Kit (Cat. No. AM1839) for gene expression analysis. I would now like to isolate miRNA. Can I still use this kit?

Yes, adjust the binding solution and wash solution 1 to 50% isopropanol to precipitate small RNAs. However, we have not yet tested yield with this protocol modification.
Are MagMax kits compatible with RNAlater reagent-treated samples?

Only MagMax96 for Microarrays Total RNA Isolation Kit (Cat. No. AM1839) is compatible, since it uses Trizol reagent and most of the salt carry-over from RNAlater reagent is eliminated during the lysis steps and partitioned from the aqueous RNA layer on top. For other MagMax kits, optimization is needed. High salt carry over is not optimal for MagMax RNA isolation steps.
I am getting MagMAX bead carryover in some wells of my plate. What might be causing this and how can I resolve it?

Some bead carryover is common and has not been documented to affect downstream assays. The most common cause of bead carryover is too much input sample. The best way to determine if the sample amount is the cause of the issue is to run PBS with Xeno (artificial DNA/RNA) in the lysis buffer and then check if there is still bead carryover. If that is not the cause, the initial sample may require special processing, such as bead beating or liquification. If there is still bead carryover without the original sample, and in particular, if the carryover is present in the same wells for multiple runs, then this may be an instrument alignment issue.
Do I need any additional consumables when using MagMAX kits on the MagMAX Express-96 instrument or a KingFisher instrument?

Plastic consumables need to be purchased separately in order to use the MagMAX reagents on the MagMAX Express-96 or a KingFisher instrument. The particular consumables and the amount required will vary by kit and protocol so please check the user guide for the particular MagMAX kit to be sure that all required plastic consumables are available prior to sample processing.

Where can I find Bindlt script protocol (.bdz) files for use of MagMAX kits with the KingFisher Flex Purification System and KingFisher Duo Prime Purification System?

The best place to download KingFisher Flex and KingFisher Duo Prime protocols for a particular MagMAX kit is the relevant MagMAX kit product page. The Bindlt script protocol (.bdz) files can be found under the Documents>Product literature section.

Can I scale up the MagMAX kit reaction volumes? If so, how should I do this?

Yes, reaction volumes for MagMAX kits can be scaled up. In general, the amount of MagMAX reagents should be scaled up proportionally for the increase in sample input amount. Please see the specific kit user guide for any recommendation. If none are included, then optimization may be necessary.

Will addition of BME (2-mercaptoethanol) to the MagMAX lysis buffer cause any issues with nucleic acid extraction?

Addition of BME (2-mercaptoethanol) to the lysis buffer of MagMAX kits is fine. This can be beneficial with more RNase-rich samples like certain types of tissues.
In general, we recommend adding 0.3 µL BME per 100 µL of lysate.

I want to use the MagMAX-96 for Microarrays Total RNA Isolation Kit to isolate total RNA with the modified spin protocol (https://bit.ly/2Z9cPQK). Can I add the TURBO DNase steps from the standard no-spin protocol to the modified spin protocol?

When using the MagMAX-96 for Microarrays Total RNA Isolation Kit modified spin protocol (https://www.thermofisher.com/us/en/home/references/protocols/nucleic-acid-purification-and-analysis/rna-protocol/isolating-small-rnaas-using-the-magmax-96-for-microarrays-kit.html) for isolating large + small RNA, we recommend going through the RNA isolation procedure and then treating the eluted RNA with TURBO DNase.
Is the MagMAX-96 for Microarrays Total RNA Isolation Kit (Cat. No. AM1839) compatible with samples treated with RNAlater Stabilization Solution (Cat. No. AM7021)?

Yes, MagMAX-96 for Microarrays Total RNA Isolation Kit is compatible with the RNAlater solution, because the kit uses Tri-Reagent. Therefore, most of the salts are removed during the lysis steps and partitioned from aqueous RNA layer on top. Additionally, salt carry-over from the RNAlater solution is not an issue.

Can I edit the manufactured KingFisher Flex protocols?

No, manufactured protocols are locked for editing, but they can be renamed using the ‘save as' function and edited as a new version.
Can I use magnetic beads from other vendors with the KingFisher Flex instrument?

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.

I am working with liquid samples with high volume and want to extract nucleic acid from them using the KingFisher Flex instrument. Can I use more volume than the protocol advises?

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.

Can I scale up my reaction volume with the KingFisher Flex instrument?

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.

Are all of the Applied Biosystems MagMAX kits validated for the KingFisher Flex Magnetic Particle Processor?

Yes, all MagMAX kits can be used with the KingFisher Flex Magnetic Particle Processor.

For Research Use Only. Not for use in diagnostic procedures.