Zymo Research YeaStar RNA Kit User manual

YeaStar™RNA Kit
RNA from any tough-to-lyse sample
Highlights
• Quick, spin-column purification of total RNA (including
small/microRNAs) from a wide range of fungi species, susceptible to
yeast lytic enzyme lysis.
• Omits the use of mechanical homogenization and organic
denaturants.
• RNA is ready for Next-Gen Sequencing, RT/qPCR and any
downstream application, etc.
Catalog Numbers:
R1002
Scan with your smart-phone camera to
view the online protocol/video.

Table of Contents
Product Contents............................................ 01
Specifications.................................................. 02
Product Description........................................ 03
Protocol ........................................................... 04
(I) Buffer Preparation ................................. 04
(II) Total RNA Purification.......................... 05
Ordering Information ...................................... 06
Complete Your Workflow ............................... 07
Notes ................................................................ 08
Guarantee ........................................................ 09
INSTRUCTION MANUAL Ver.2.0.2 Revised on: 8/9/2021

1
Product Contents
YeaStar™RNA Kit R1002
(40 prep)
YR Digestion Buffer 3.2 ml
YR Lysis Buffer 6.4 ml
RNA Wash Buffer
1
(concentrate) 6 ml
DNase/RNase-Free Water 6 ml
Zymolyase
2
(lyophilized) 1000 U
Zymolyase Storage Buffer 0.5 ml
Zymo-Spin™IIICG Columns 50
Collection Tubes 50
Instruction Manual 1
Storage Temperature - Store all kit components (i.e., buffers, columns) at room temperature.
Before use:
1 Add 24 ml 100% ethanol (26 ml 95% ethanol) to the 6 ml RNA Wash Buffer concentrate.
2 Add 200 µl Zymolyase Storage Buffer to the lyophilized Zymolyase and mix by gentle inversion. Store frozen
aliquots.

2
Specifications
• Sample Sources – Any fungi species (1-5 x 107pelleted cells
from 1-1.5 ml culture), susceptible to yeast lytic enzyme lysis.
RNA can also be isolated from aged cells grown on solid or in
liquid medium. Specific sample types include, but are not limited
to: Aspergillus fumigatus, Aspergillus nidulans, Aspergillus nivens
var. aureus, Candida albicans, Pichia pastoris, Saccharomyces
cerevisiae, Schizosaccharomyces pombe.
• Size – Total RNA including small/microRNAs (≥ 17 nt).
• Purity – A260/A280 & A260/A230 > 1.8. RNA is ready for Next-Gen
Sequencing, RT/qPCR, etc.
• Binding Capacity – Zymo-Spin™IIICG Column yield up to
100 µg RNA.
• Elution Volume – ≥ 35 µl DNase/RNase-Free Water.
•Equipment Needed (user provided) – Microcentrifuge, vortex.

3
Product Description
The YeaStar™RNA Kit provides for rapid isolation of total RNA from a
wide range of fungi species, susceptible to yeast lytic enzyme lysis
including Aspergillus fumigatus, Aspergillus nidulans, Aspergillus nivens
var. aureus, Candida albicans, Pichia pastoris, Saccharomyces
cerevisiae, Schizosaccharomyces pombe.
The Zymo-Spin™IIICG Column efficiently binds total RNA and is eluted
in ≥ 35 µl water, suitable for subsequent procedures including RT-qPCR.

4
Protocol
The protocol consists of: (I) Buffer Preparation and (II) Total RNA
Purification
(I) Buffer Preparation
Add 24 ml 100% ethanol (26 ml 95% ethanol) to the 6 ml RNA Wash
Buffer.
Add 200 µl Zymolyase Storage Buffer to the lyophilized Zymolyase
(final concentration 5 U/µl) and mix by gentle inversion. Store frozen
aliquots.

5
(II) Total RNA Purification
Perform all steps at room temperature and centrifugation at 10,000-16,000 x g for 30
seconds, unless specified.
1. Pellet 1-5 x 107cells (1-1.5 ml culture) by centrifugation at 500 x g for
2 minutes. Then carefully remove the supernatant.
2. Add 80 µl of the YR Digestion Buffer and 5 µl of the Zymolyase1to
the cell pellet and resuspend the pellet completely by pipetting up and
down.
3. Incubate the suspension at 30-37°C for 40-60 minutes2.
If the volume of the cell pellet is ≤ 25 µl, incubate for 40 minutes. For cell pellet volume
≥ 25 µl, incubate for 60 minutes.
4. Add 160 µl of the YR Lysis Buffer and mix thoroughly by vortexing.
5. Add an equal volume ethanol (95-100%) (1:1) and mix well.
6. Transfer the mixture to the Zymo-Spin™IIICG Column3in a
Collection Tube and centrifuge. Discard the flow-through.
7. Add 200 µl RNA Wash Buffer to the column and centrifuge. Discard
the flow-through.
8. Add 200 µl RNA Wash Buffer to the column and centrifuge. Then
carefully, transfer the column into a nuclease-free tube (not provided).
9. Add 60 µl of DNase/RNase-Free Water directly to the column matrix
and centrifuge
Alternatively, for highly concentrated RNA use ≥ 35 µl elution.
The eluted RNA can be used immediately or stored frozen.
1 Prior to use, resuspend the lyophilized Zymolyase according to Buffer Preparation, page 4.
2. Incubation time will vary based on cell number. Optimization may be required.
3 To process samples > 700 µl, columns may be reloaded.

6
Ordering Information
Product Description Catalog No. Size
YeaStar™RNA Kit R1002 40 preps.
Individual Kit Components Catalog No. Amount
YR Digestion Buffer R1001-1 3.2 ml
YR Lysis Buffer R1001-2 6.4 ml
RNA Wash Buffer (concentrate) R1003-3-6
R1003-3-24
6 ml
24 ml
DNase/RNase-Free Water W1001-10
W1001-30
10 ml
30 ml
Zymolyase (lyophilized; with Storage Buffer)
E1004
E1005
E1006 (w/ RNase A)
1000 U
2000 U
1000 U
Zymo-Spin™IIICG Columns C1006-50-G 50
Collection Tubes C1001-50 50

7
Complete Your Workflow
For tough-to-lyse samples, use ZR BashingBead Lysis Tubes:
ZR BashingBead Lysis Tubes
2.0 mm beads #S6003 Plant/animal tissue
0.1 + 0.5 mm beads #S6012 Microbes
0.1 + 2.0 mm beads #S6014 Microbes in tissue/insects
For isolation of RNA from any sample:
Quick-RNA Fungal/Bacterial Kits
Microprep #R2010
Includes ZR BashingBead
Lysis Tubes for mechanical
homogenization of tough-to-lyse
microbial samples.
Miniprep #R2010
For clean-up (purification) and concentration of any RNA sample. (e.g., from the
aqueous phase of TRIzol®extractions) or from any enzymatic reaction (e.g., DNase I
treated RNA):
RNA Clean & Concentrator kits
Microprep #R1013-R1014 DNase I Set included
MagBeads #R1082
Automatable (Tecan, Hamilton,
Kingfisher, etc.)
For NGS:
Zymo-Seq RiboFree Total RNA Library Prep kit
#R3000 12 preps
#R3003 96 preps

8
Notes

9
100% satisfaction guarantee on all Zymo Research products,
or your money back.
Zymo Research is committed to simplifying your research with quality products
and services. If you are dissatisfied with this product for any reason, please call
1(888) 882-9682.
Integrity of kit components is guaranteed for up to one year from date of purchase.
Reagents are routinely tested on a lot-to-lot basis to ensure they provide the highest performance and reliability.
This product is for research use only and should only be used by trained professionals. It is not for use in
diagnostic procedures. Some reagents included with this kit are irritants. Wear protective gloves and eye
protection. Follow the safety guidelines and rules enacted by your research institution or facility.
™ Trademarks of Zymo Research Corporation
YeaStar®is a registered trademark of Zymo Research Corporation.