Zymo Research는 1994년도에 설립된 미국 회사로 고품질 키트 제조회사 입니다.
각종 샘플(세포, 조직, 환경샘플 등)로부터 고품질
DNA나 RNA를 가장 쉽고 빠르게 뽑을 수 있습니다.
이 외에도, 각종 Epigenetics 관련 제품들
(DNA Methylation kit 등)과 Microbiomics
(샘플 채집부터 분석까지의 전 단계의 제품)
연관 제품들이 준비되어 있습니다.
* 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
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 Input
200 ng of total RNA, or RNA extracted from 200 µL of plasma or serum.
Minimum Input
1 ng of total RNA, or RNA extracted from 50 µL of plasma or serum.
Processing Time
2 hours hands-on, 8 hours total
Recommended Equipment
A 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 Input
100 ng of total RNA, or RNA extracted from 200 µL of plasma or serum.
Required Equipment
Thermal cycler, magnetic stand for 0.2 mL PCR tubes or 96-well plate, mini centrifuge, and micro centrifuge.
Sample Type
High 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 Compatibility
Libraries are compatible with all Illumina sequencing platforms.
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 Microprep, Quick-RNA Microprep, and Quick-cf-RNA Serum & Plasma Kit.
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.
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.
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.
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.
