TITLE: ZymoBIOMICS MagBead DNA/RNA-R2135
AUTHORS: ZYMO Research Corp.

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">The ZymoBIOMICSTM MagBead DNA/RNA kit provides a high-throughput, magnetic bead-based purification of both high-quality DNA and total RNA (including small/microRNAs) from the same starting sample. The provided DNA/RNA ShieldTM inactivates infectious agents and is ideal for sample storage at ambient temperatures. The extraction method utilizes magnetic beads for DNA/RNA extraction without the use of phenol and is eluted into ≥50 μl of ZymoBIOMICSTM DNase/RNase-Free Water. DNA/RNA is ready for any downstream application including Next-Gen Sequencing, RT-qPCR, etc.</div><div class = "text-block">Unbiased Lysis: Efficient and unbiased lysis of microbes including gram positive/negative bacteria, fungi, protozoans, and viruses from any sample including feces, soil, plant, water, biofilms, swabs, saliva, body fluids, etc.</div><div class = "text-block">• Ultra-Pure: High-quality DNA/RNA (including small/microRNAs) are inhibitor-free and ready for RT/qPCR and microbiome measurements using Next-Gen sequencing.</div><div class = "text-block">• High-Sensitivity: Increased detection limit of very low abundance organisms.</div></div>

[STEPS]
?. [Sample Preparation]
Add to a sample 250 uL max.input and mix and/or homogenize.  AB1Sample TypeMaximum Input2Feces50 mg3Soil50 mg4Liquid Samples and Swab Collections250 ul5Cells (suspended in DNA/RNA Shield or isotonic buffer, e.g. PBS)5-20 mg (wet weight; 2x10^8 bacterial, 2x10^7 yeast cells, 2x10^6 mammalian cells)6Samples in DNA/RNA Shield (10% v/v)250 ul
All centrifugation steps should be performed at 10,000 - 16,000 x g for 30 seconds, unless specified.For 96-well lysis rack, centrifuge at 4,000 x g for 5 minutes.All steps should be performed at room temperature (20-30oC), unless specified.
[DNA/RNA Shield]
AB1Sample TypeMaximum Input2Feces50 mg3Soil50 mg4Liquid Samples and Swab Collections250 ul5Cells (suspended in DNA/RNA Shield or isotonic buffer, e.g. PBS)5-20 mg (wet weight; 2x10^8 bacterial, 2x10^7 yeast cells, 2x10^6 mammalian cells)6Samples in DNA/RNA Shield (10% v/v)250 ul
At this point, samples in DNA/RNA Shield can be stored at ambient temperature (4-25°C) for a month, 3 days at 37°C, or long-term (> 1 year) -20°C or below.
?. [Total Nucleic Acid Purification]
Add to 200 ul sample and mix well.
[DNA/RNA Lysis Buffer]
?. [DNA and RNA Purification]
Add to the 200 ul sample and mix well.
[DNA/RNA Lysis Buffer]
?. [Total Nucleic Acid Purification]
Add to the sample and mix well.
[ethanol (95-100%)]
?. [Total Nucleic Acid Purification]
Add and mix well for 20 minutes.
[ZymoBIOMICS MagBinding Beads]
Important: ZymoBIOMICS MagBinding Beads settle quickly, ensure that beads are kept in suspension while dispensing.
?. [Total Nucleic Acid Purification]
Repeat once.
?. [Total Nucleic Acid Purification]
DNase I treatment (optional)
?. [DNA and RNA Purification]
Add and mix well for 20 minutes.
[ZymoBIOMICS MagBinding Beads]
Important: ZymoBIOMICS MagBinding Beads settle quickly, ensure that beads are kept in suspension while dispensing.
?. [DNA and RNA Purification]
Transfer the plate/tube to the magnetic stand until beads (DNA) have pelleted, then transfer the cleared supernatant (RNA) into a new plate/tube.
?. [Sample Preparation]
To achieve unbiased lysis of different organisms, including hard-to-lyse microbes, perform mechanical homogenization3 (Recommended e.g., lysis tubes S6012-50 or lysis rack S6002-96-3; each sold separately). Then centrifuge to pellet debris and transfer 200 μl supernatant to a new tube.
?. [Sample Preparation]
Add 10 μl Proteinase K for every 200 μl sample. Mix and incubate at room temperature (20-30oC) for 30 minutes.
?. [Sample Preparation]
Proceed to Total Nucleic Acid Purification, page 4 or DNA and RNA Purification, page 5.
?. [Total Nucleic Acid Purification]
Transfer the plate/tube to the magnetic stand until beads have pelleted, then aspirate and discard the cleared supernatant.
?. [Total Nucleic Acid Purification]
Add and mix well. Pellet the beads discard the supernatant.
[MagBead DNA/RNA Wash 1]
?. [Total Nucleic Acid Purification]
Add and mix well. Pellet the beads discard the supernatant.
[MagBead DNA/RNA Wash 2]
?. [Total Nucleic Acid Purification]
Add and mix well. Pellet the beads discard the supernatant.
[ethanol (95-100%)]
?. [Total Nucleic Acid Purification]
Dry the beads for 10 minutes or until dry.
?. [Total Nucleic Acid Purification]
Add 50 μl DNase I Reaction Mix and mix gently for 10 minutes.
?. [Total Nucleic Acid Purification]
Add 500 μl DNA/RNA Prep Buffer and mix well1 for 10 minutes. Pellet the beads2,3 and discard the supernatant.
?. [Total Nucleic Acid Purification]
Repeat once.
?. [Total Nucleic Acid Purification]
To elute DNA/RNA from the beads, add 50 μl ZymoBIOMICS DNase/RNase- Free Water and mix well1 for 5 minutes.
?. [Total Nucleic Acid Purification]
Transfer the plate/tube to the magnetic stand until beads have pelleted, then aspirate and dispense the eluted DNA/RNA to a new plate/tube.
?. [DNA and RNA Purification]
Add and mix well. Pellet the beads and discard the supermatant.
[MagBead DNA/RNA Wash 1]
?. [DNA and RNA Purification]
Repeat once.
?. [DNA and RNA Purification]
Add and mix well. Pellet the beads and discard the supermatant.
[MagBead DNA/RNA Wash 2]
?. [DNA and RNA Purification]
Add and mix well. Pellet the beads and discard the supermatant.
[ethanol (95-100%)]
?. [DNA and RNA Purification]
Dry the beads for 10 minutes or until dry.
?. [DNA and RNA Purification]
Add and mix well for 5 minutes.
[ZymoBIOMICS DNase/RNase-Free Water]
?. [DNA and RNA Purification]
Transfer the plate/tube to the magnetic stand until beads have pelleted, then aspirate and dispense the eluted DNA to a new plate/tube.
?. [DNA and RNA Purification]
Add to the supermatant and mix well.
[ethanol (95-100%)]
?. [DNA and RNA Purification]
Add and mix well for 10 minutes.
[/well ZymoBIOMICS MagBinding Beads]
?. [DNA and RNA Purification]
Transfer the plate/tube to the magnetic stand until beads (DNA) have pelleted, then transfer the cleared supernatant (RNA) into a new plate/tube.
?. [DNA and RNA Purification]
Add and mix well. Pellet the beads discard the supernatant.
[MagBead DNA/RNA Wash 1]
?. [DNA and RNA Purification]
Add and mix well. Pellet the beads discard the supernatant.
[MagBead DNA/RNA Wash 2]
?. [DNA and RNA Purification]
Add and mix well. Pellet the beads and discard the supermatant.
[ethanol (95-100%)]
?. [DNA and RNA Purification]
Repeat once.
?. [DNA and RNA Purification]
DNase I treatment (optional)
?. [DNA and RNA Purification]
Add 50 μl DNase I Reaction Mix and mix gently for 10 minutes.
?. [DNA and RNA Purification]
Add 500 μl DNA/RNA Prep Buffer and mix well1 for 10 minutes. Pellet the beads2,3 and discard the supernatant.
?. [DNA and RNA Purification]
Repeat once.
?. [DNA and RNA Purification]
Dry the beads for 10 minutes or until dry.
?. [DNA and RNA Purification]
Add and mix well for 5 minutes.
[ZymoBIOMICS DNase/RNase-Free Water]
?. [DNA and RNA Purification]
Transfer the plate/tube to the magnetic stand until beads have pelleted, then aspirate and dispense the eluted DNA to a new plate/tube.