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Invitrogen™ MagMAX™ for Stabilized Blood Tubes RNA Isolation Kit, compatible with Tempus™ Blood RNA Tubes

Catalog No. 4451893
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Catalog No. 4451893 Supplier Invitrogen™ Supplier No. 4451893
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Description

Includes

Kit Components Box 1: Tempus™ Resuspension Solution, 12mL; Binding Solution Concentrate 5mL; Wash Solution 1 Concentrate, 2 x 16mL (Before using the Wash Solution 1 Concentrate, add the correct volume of isopropanol (user-supplied), as indicated on the bottle); Wash Solution 2 Concentrate, 30mL (Before using the Wash Solution 2 Concentrate, add the correct volume of ethanol (user-supplied), as indicated on the bottle); Elution Buffer 10mL; Processing Plate (96-well) 1 plate, Tempus™ Pre-Digestion Wash, 2 x 200mL; RNA Binding Beads, 2mL; Tempus™ 1X PBS, 2 x 150mL. Box 2: Tempus™ Proteinase, 0.25mL; TURBO™ DNase, 1mL

Requires

Refrigerated centrifuge (4°C) with swing bucket rotor capable of 5,000 x g; 50mL conical tube adapters; 50mL conical tubes; Tabletop microcentrifuge or picocentrifuge (for For tube format users); Blood tubes; Magnetic stand

The MagMAX for Stabilized Blood Tubes RNA Isolation Kit purifies total RNA (including miRNA) from Tempus Blood RNA Tubes using proven magnetic bead-based technology.

The MagMAX for Stabilized Blood Tubes RNA Isolation Kit purifies total RNA (including miRNA) from Tempus Blood RNA Tubes using proven magnetic bead-based technology. A TURBO DNase™ step efficiently eliminates contaminating gDNA ensuring the highest purity RNA possible suitable for real-time RT-PCR and microarray analysis.

Maximize Total RNA Yields From Blood Samples
The MagMAX for Stabilized Blood Tubes RNA Isolation Kit utilizes proven magnetic bead-based technology to purify maximal yields of total RNA from Tempus™ Blood RNA Tubes. The MagMAX for Stabilized Blood Tubes RNA Isolation Kit can recover 3-25 μg from a single Tempus Blood RNA tube. A TURBO DNase step efficiently eliminates contaminating gDNA that can interfere with downstream applications and ensures the highest purity RNA possible. The MagMAX for Stabilized Blood Tubes RNA Isolation Kit can also detect miRNA from Tempus Blood RNA tubes as shown by TaqMan™ gene expression assays. Users are strongly advised to perform appropriate validation studies against their miRNA targets.

Rapid Processing Time
Processing times will vary based on format selected, but twelve Tempus Blood RNA tubes can be processed manually with the MagMAX for Stabilized Blood Tubes RNA Isolation Kit in approximately 2 hours.

An Alternative to Full Purification
For users who perform primarily TaqMan-based real-time RT-PCR experiments, and do not need to archive RNA/cDNA, an alternative to the MagMAX for Stabilized Blood Tubes RNA Isolation Kit is the Stabilized Blood-to-CT™ Nucleic Acid Preparation Kit for qPCR. This kit makes it possible to perform expression analysis directly from Tempus Blood RNA tubes without conventional RNA purification.

One-Stop Shopping
MagMAX for Stabilized Blood Tubes RNA Isolation Kit is compatible with reverse transcription mixes (SuperScript™), PCR master mixes (both TaqMan and SYBRbased), and other blood workflow products.

  • Flexible purification format—use microfuge tubes, manual 96-well plates, or open-format automated 96-well plates from the same kit
  • Simple magnetic bead workflow—no spin columns
  • Total RNA without RNase activity, contaminating gDNA, or RT-PCR inhibitors
  • Suitable for use with automated protocols, high-throughput compatible

Flexible Purification Formats

  • The MagMAX for Stabilized Blood Tubes RNA Isolation Kit can be processed in low-, medium- or high-throughput formats with superior workflow and handling compared to traditional competitive methods
  • For low-throughput users, the MagMAX for Stabilized Blood Tubes RNA Isolation Kit can be used in individual 1.5 mL microfuge tubes with the bead-RNA-complex remaining in a single processing tube until the final elution step
  • For medium-to-high throughput users, the MagMAX for Stabilized Blood Tubes RNA Isolation Kit can be done in a 96-well processing plate with either a single or multi-channel pipettor to accelerate the purification process
  • For high-throughput users, the MagMAX for Stabilized Blood Tubes RNA Isolation Kit is compatible with manual and automated 96-well processing
  • Simply process multiple batches of tubes in series or in parallel, through the up-front centrifugation process, to the convenient stopping point, until enough samples can be loaded onto a 96-well processing plate for the RNA purification steps
  • The RNA purification process can be handled on either open-format liquid handlers, in conjunction with a 96-well magnetic stand on the deck, or magnetic bead movers like the MagMAX Express-96 Magnetic Particle Processor (deep well version only)

Specifications

Content And Storage

Box 1
• 12 mL Tempus Resuspension Solution; room temperature
• 5 mL Binding Solution (concentrated); room temperature
• 2 x 16 mL Wash Solution 1 (concentrated); room temperature
• 30 mL Wash Solution 2 (concentrated); room temperature
• 10 mL Elution Buffer; room temperature
• 1 Processing Plate; room temperature
• 2 x 200 mL Tempus Pre-Digestion Wash; 4°C
• 2 mL RNA Binding Beads; 4°C
• 2 x 150 mL Tempus 1X PBS; 4°C

Box 2
• 0.25 mL Tempus Proteinase; -20°C
• 1 mL TURBO DNase; -20°C
Isolation Technology Magnetic Bead
Purification Time Less than 2 hr. for automated protocol
Elution Volume 20 to 80 μL
Sample Type Whole Blood
Final Product Type Total RNA
High-throughput Compatibility High-throughput Compatible
For Use With (Equipment) KingFisher™ Duo Prime, KingFisher™ Flex
For Use With (Application) RT-PCR, qPCR, cDNA library construction, NGS, microarray analysis, blot hybridization, Northern blotting, in vitro translation, nuclease protection assays, nucleic acid labeling
Quantity 96 preps
Shipping Condition Box 1: Room Temperature
Box 2: Cold Gel
Starting Material Amount 3 mL of whole blood collected in Tempus™ Blood RNA Tube
Yield ≤25 μg
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Frequently Asked Questions (FAQs)

I am doing blood work and using the Tempus collection tubes. Is the Tempus system compatible with the RiboPure-Blood RNA Purification Kit?

No, please follow the RNA isolation methods described with the Tempus system. The collection tubes are not compatible with the RiboPure kit.
Do you have any tips for RNA isolation when working with blood samples?

Please visit our website for tips for working with blood samples.
Can I purchase the RNA Binding Beads from the MagMAX for Stabilized Blood Tubes RNA Isolation Kit, compatible with Tempus Blood RNA Tubes (Cat. No. 4451893) separately?

No, the RNA Binding Beads are not available as a stand-alone product.
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.

Is there a good stopping point in the workflow for the MagMAX for Stabilized Blood Tubes RNA Isolation Kit, compatible with Tempus Blood RNA Tubes (Cat. No. 4451893)?

The user guide for our MagMAX for Stabilized Blood Tubes RNA Isolation Kit, compatible with Tempus Blood RNA Tubes, states that you can stop after the RNA resuspension step by storing the resuspended crude RNA pellet on ice for up to 4 hours.
Please see page 18 of the following user guide: https://assets.thermofisher.com/TFS-Assets/LSG/manuals/cms_084749.pdf.
Can animal blood stored in Tempus Blood RNA Tubes or PAXgene tubes be prepared with the MagMAX for Stabilized Blood Tubes RNA Isolation Kits (Cat. Nos. 4451893 & 4451894)?

Equine whole blood has been successfully tested using this system, getting RINs around 9. Bovine blood should work similarly since there is not much of a difference between the two species for RNA isolation from whole blood, but this has not been tested. The protocol would be the same as the user guide and samples should be mixed vigorously immediately after blood is drawn into the tube for at least 10 secs, incorporating tube inversion to be sure the sample is stabilized.
Is the MagMAX mirVana Total RNA Isolation Kit (Cat. No. A27828) compatible with Tempus/PAXgene stabilized blood samples?

No, only certain MagMAX kits are compatible with Tempus/PAXgene stabilized blood samples. We recommend using only MagMAX kits that are designed for blood samples stored in the designated RNA blood tubes for the kit. For PAXgene tubes, the compatible kit is the MagMAX for Stabilized Blood Tubes RNA Isolation Kit, compatible with PAXgene Blood RNA Tubes (Cat. No. 4451894). For Tempus tubes the compatible kit is the MagMAX for Stabilized Blood Tubes RNA Isolation Kit, compatible with Tempus Blood RNA Tubes (Cat. No. 4451893).
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.