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Zymo Research
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Zymo-Seq miRNA Library Kit
* ligation 편향성은 줄이고, cfRNA 및 total RNA의 정확한 RNA 프로파일링을 제공
Zymo Research의 혁신적인 RNA Sequencing Library Prep Kit 입니다.
홍기환 대리

Zymo Research 제품 담당자

경신과학(주)

영업부

제품소개

*이 제품은 25.5.7 부로 단종되었습니다.*
*DISCONTINUED PRODUCTS*


HIGHLIGHTS

  • Accurate miRNA profiling: A unique single adapter and circularization strategy reduces ligation bias and increases miRNA detection.
  • Direct detection: Generates small RNA libraries without the need for small RNA enrichment.
  • Gel-Free: Removes adapter dimers from any input without lengthy gel excision.
DESCRIPTION

The Zymo-Seq™ miRNA Library Kit is an innovative small RNA sequencing library prep kit that uses a single-adapter and circularization strategy to reduce ligation bias and provide accurate small RNA profiling from cell-free RNA (cfRNA) and total RNA. This simple, streamlined workflow minimizes hands-on time and uses gel-free removal of adapter dimers to generate stranded small RNA libraries from plasma, biofluids, cells, tissues, and more.

To prepare Zymo-Seq™ miRNA libraries, RNA is ligated to the single miRNA adapter and the ligated product is circularized. Excess adapter is blocked and removed with specialized magnetic beads that allow a completely gel-free workflow for input as low as 1 ng. The circularized miRNA-adapter product is then reverse transcribed. The resulting cDNA is amplified using Indexing PCR primers which add on Illumina-compatible adapter and unique dual index (UDI) sequences for cost-effective multiplexing. Everything needed to make sequencing-ready libraries, including indexing primers and SPRI beads, are included in the kit. It’s RNA-Seq made simple.


Additional Components Required (Not Provided)Molecular biology grade pure ethanol (> 95%)
Maximum Input200 ng of total RNA, or RNA extracted from 200 µL of plasma or serum.
Minimum Input1 ng of total RNA, or RNA extracted from 50 µL of plasma or serum.
Processing Time2 hours hands-on, 8 hours total
Recommended EquipmentA 96-well Aluminum Cooler Block for PCR tubes or a 96-well PCR tube rack may be placed on ice for ease of reagent assembly and maintaining samples at 4°C. To process > 12 samples at once, a multichannel pipette with low-retention tips is recommended for mixing and the timely addition of pre-aliquoted reagents.
Recommended Input100 ng of total RNA, or RNA extracted from 200 µL of plasma or serum.
Required EquipmentThermal cycler, magnetic stand for 0.2 mL PCR tubes or 96-well plate, mini centrifuge, and micro centrifuge.
Sample TypeHigh integrity total RNA (RIN > 7) containing miRNA, or cell-free RNA extracted from plasma/serum samples not preserved with heparin. Small RNA enrichment is not necessary.
Sequencing Platform CompatibilityLibraries are compatible with all Illumina sequencing platforms.



Q1: What RNA samples can I process with the Zymo-seq miRNA Library Kit?

The Zymo-seq miRNA Library Kit can be used with a variety of RNA samples that contain microRNA, including plasma, urine, saliva, exosomes, and total RNA extracted from various cells and tissues. Many purification methods do not adequately purify small RNA. It is critical that the RNA purification method used effectively retains the microRNA and small RNAs, such as Direct-zol MicroprepQuick-RNA Microprep, and Quick-cf-RNA Serum & Plasma Kit.

Q2: Can I use samples with low RNA quality?

When using total RNA, we recommend using high quality RNA with a RNA Integrity Number (RIN) value of at least 7. RNA with a lower RIN value may be used but may result in over-representation of rRNA sequences due to the presence of higher levels of fragmented rRNA. This could reduce the representation of miRNAs and other small RNAs in the final library.

Q3: Do I need to perform size selection on total RNA to enrich for small RNA before preparing libraries?

The total RNA may be used without size selection. RNAs larger than ~100 nt within the sample are not effectively incorporated into the final library.

Q4: Which sequencing platforms is the Zymo-Set miRNA Library Kit compatible with?

Zymo-Seq miRNA libraries are compatible with any Illumina® sequencer, as well as the Element Biosciences AVITI™ system (with Element Adept Library Compatibility workflow.). This kit has not been validated on Ion Torrent®, Oxford Nanopore®, PacBio®, or other non-Illumina sequencing platforms.

Q5: What read length should I sequence libraries generated using the Zymo-seq miRNA Library Kit?

Libraries may be sequenced at any length. However, it is important to note that reads longer than the typical insert size (~21nt-100nt) will require more adapter trimming through the excess sequencing cycles. For optimal results, we recommend sequencing with a minimum read length of 50 base pairs (bp) and a maximum of 75 bp and single-end sequencing.

Q6: Is technical support available for the Zymo-seq miRNA Library Kit?

We are happy to offer additional recommendations, troubleshooting assistance, and consultation requests. Email us at tech@zymoresearch.com if you require further guidance or have any questions regarding the Zymo-seq miRNA Library Kit.


 

주문정보

CAT.No 품명 규격 비고
R3006 Zymo-Seq miRNA Library Kit 12 preps
R3007 Zymo-Seq miRNA Library Kit 96 preps