![[NEB] 신제품 NEBNext® High Input Poly(A) mRNA Isolation Module](https://www.korambiotech.com/wp-content/uploads/2023/03/230327-NEBNext®-High-Input-PolyA-mRNA-Isolation-Module-큰-1.jpg)
NEBNext® High Input Poly(A) mRNA Isolation Module은 oligo d(T)25-coupled paramagnetic beads를 사용하여 많은 양의 total RNA (5-50 µg per 0.2 ml reaction)에서 intact mRNA를 한 시간 내에 얻을 수 있는 제품입니다.
5 µg이하의 total RNA를 사용할 경우에는 기존 제품인 NEBNext® Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) 사용을 권장합니다.
![[NEB] 신제품 NEBNext® High Input Poly(A) mRNA Isolation Module](https://www.korambiotech.com/wp-content/uploads/2023/03/1.jpg)
Figure 1. The NEBNext High Input Poly(A) mRNA Isolation Module provides consistent representation across a wide input range
![[NEB] 신제품 NEBNext® High Input Poly(A) mRNA Isolation Module](https://www.korambiotech.com/wp-content/uploads/2023/03/1-1.jpg)
A. Poly(A) RNA enriched using the NEBNext High Input Poly(A) mRNA Isolation module from 50 μg or 5 μg of total Universal Human Reference (UHR) RNA (Agilent®) containing External RNA Controls Consortium (ERCC) RNA and Spike-In RNA Variants (SIRV Set 4, Lexogen®). Total yield was assessed using Qubit® RNA High Sensitivity Assay Kit (Invitrogen®). Poly(A) RNA is typically ~1-5% of total RNA.
B. Bioanalyzer® (Agilent) traces for Total UHR RNA and poly(A)-enriched RNA from 50 μg or 5 μg total RNA input.
C. Transcript abundances were determined from libraries prepared from 40 ng (50 μg input) or 8 ng (5 μg input) poly(A)-enriched RNA using the NEBNext Ultra™ II Directional RNA Library Prep Kit and sequenced on an Illumina® NextSeq® 550 sequencer. Six million reads were sampled from each library.
Figure 2. The NEBNext High Input Poly(A) mRNA Isolation Module produces low ribosomal RNA retention, across sample types
![[NEB] 신제품 NEBNext® High Input Poly(A) mRNA Isolation Module](https://www.korambiotech.com/wp-content/uploads/2023/03/2.jpg)
Poly(A) RNA was enriched using Dynabeads® mRNA Purification Kit (Invitrogen), Poly(A)Purist™ MAG (Invitrogen) or the NEBNext High Input Poly(A) mRNA Magnetic Isolation Module from 50 μg Universal Human Reference RNA (UHR, Agilent) or RNA extracted from mouse kidney tissue or S. cerevisiae (yeast) using the Monarch® Total RNA Miniprep Kit.
A. Representative Bioanalyzer traces of total and poly(A)-enriched RNA.
B. Percent ribosomal RNA (rRNA) of total or poly(A)-enriched RNA samples was determined from sequencing of triplicate (UHR and mouse poly(A) samples) or duplicate (total RNA and yeast poly(A) RNA samples) experiments, with standard deviation. Libraries were prepared from 40 ng poly(A)-enriched RNA using the NEBNext Ultra™ II Directional RNA Library Prep Kit and sequenced on an Illumina NextSeq 550 instrument. Six million reads were sampled from each library.
Q&A
No, this kit is intended for isolation of mRNA from previously isolated total RNA. Total RNA can be isolated using the Monarch® Total RNA Miniprep Kit (NEB #T2010).
For best results when using total RNA inputs higher than 50 µg, all volumes in the protocol can be proportionally scaled. For inputs less than 5 µg, the NEBNext® Poly(A) mRNA Magnetic Isolation Module (NEB #E7490) is recommended.
No. In order to avoid sample carryover contamination, re-use of the beads is not recommended.
Sequencing platform, sample type에 맞춰 권장하는 NEB Next generation sequencing (NGS) 제품을 선택할 수 있는 Tool입니다.
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