NEXTFLEX® Poly(A) Beads 2.0

NEXTFLEX® Poly(A) Beads 2.0

The kit offers a convenient method for the purification of pure, intact mRNA upstream of RNA-Seq library prep. Poly(A)-tailed mRNA is isolated via magnetic beads conjugated to oligo(dT), and separation using a magnetic stand allows for high mRNA recovery. The intact mRNA is eluted in small volumes, thereby eliminating the need for precipitation. These beads have been validated with the new and improved NEXTFLEX® Rapid Directional RNA-Seq Kit 2.0.

 

产品特点：

• 10 ng – 5 µg total starting RNA
• Magnetic bead-based protocol
• No organic solvents or precipitation step required
• Validated using the NEXTFLEX® Rapid Directional RNA-Seq Kit 2.0
• Magnetic bead-based protocol for purification of pure, intact mRNA upstream of RNA-Seq library prep (no organic solvents or precipitation step required)
• Optimized for use with 10 ng – 5 µg of total RNA as starting material
• Validated using the NEXTFLEX® Rapid Directional RNA-Seq Kit 2.0 and automated on the Sciclone® G3 NGSx and Zephyr® G3 NGS workstations

 

产品列表：

货号	产品名称	规格
NOVA-512991	NEXTFLEX® Poly(A) Beads 2.0	8 RXNS
NOVA-512992	NEXTFLEX® Poly(A) Beads 2.0	48 RXNS
NOVA-512993	NEXTFLEX® Poly(A) Beads 2.0	96 RXNS

NEXTFLEX® Poly(A) Beads 2.0 for Reducing Ribosomal rRNA Reads in RNA-seq Libraries 

Several approaches have been used to remove ribosomal RNA (rRNA) from total RNA samples for RNA-Seq library preparation. Removal of rRNA avoids the waste of reagents and bioinformatics resources to sequence and align ~85% of total RNA comprising rRNA (which is generally not a target of interest in NGS sequencing experiments). PerkinElmer’s NEXTFLEX® Poly(A) beads 2.0 kit offers an easy and cost-effective method for removing rRNA in RNA-Seq libraries. This product takes advantage of the tract of adenosine residues present at the 3’ends of a vast majority of protein-coding mRNAs. Hybridization of these poly(A) tails to tracts of complementary thymidines (“oligo dT”) immobilized on solid supports allows the retrieval of poly(A)+ RNAs by recovering the solid support along with the hybridized RNA complexes.

Figure 1.Example of 1 µg of Universal Human Reference RNA (Agilent® # 740000) after Poly(A) enrichment using the NEXTFLEX® Poly(A) Beads 2.0.  3 µL Poly(A) enriched RNA was run on the LabChip® GXII Touch™ HT instrument using the RNA Pico Assay Reagent Kit (# CLS960012) and a DNA/RNA/Charge Variant Assay LabChip (# 760435)

Figure 2. NEXTFLEX® Poly(A) Beads 2.0 yield clean mRNA with minimal rRNA carry-over from 5 µg of total RNA input from (A) mouse brain and (B) MCF7 cells. ~1.5% rRNA carry-over was observed based on sequencing data.

试剂组分：

NEXTFLEX® POLY(A) BEADS KIT 2.0 CONTENTS

• NEXTFLEX® Poly(A) Beads 2.0
• NEXTFLEX® Poly(A) Elution Buffer 2.0
• NEXTFLEX® Poly(A) Binding Buffer 2.0
• NEXTFLEX® Poly(A) Washing Buffer 2.0

REQUIRED  MATERIALS NOT PROVIDED

• 10 ng -5 μg total RNA
• DynaMag™-2 Magnet (Life Technologies Cat # 123-21D) / or / similar
• Water bath / Heat blocks
• Tube Rotator-unit (VWR Cat # 13916-822) / or / similar
• 2, 10, 20, 200 and 1000 μL pipettes
• RNase-free pipette tips
• Nuclease-free test tubes and microcentrifuge tubes
• Microcentrifuge
• Vortex

技术服务：
1、DAP-seq：高通量检测转录因子或DNA结合蛋白在基因组上的结合位点。
2、酵母单杂交：研究蛋白质和特定DNA序列相互作用。
3、酵母双杂交：检测已知蛋白质之间的相互作用，鉴定与已知蛋白相互作用的未知蛋白。
4、EMSA：鉴定或验证DNA结合蛋白和DNA序列的相互作用。
5、ChIP-seq：高效检测重组蛋白、转录因子在基因组的结合位点。
6、DAP-seq与RNA-seq联合分析： 分析转录因子的靶基因在RNA-seq数据中的表达变化，深入挖掘DAP-seq和RNA-seq测序数据，增加转录组测序的分析深度。
7、DNA-pull down：鉴定与DNA结合的蛋白。
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