NEXTFLEX 16S V4 and 18S Amplicon-Seq Kit

Ideal for species identification of bacteria, fungal, and micro-eukaryotic organisms from uncultured samples of complex communities

The NEXTFLEX® 16S V4 and 18S Amplicon-Seq Kit is designed to prepare libraries that cover both the V4 hypervariable domain of the 16S rRNA genes present in microbial populations and the 18S regions that can identify fungal and micro-eukaryotic organisms in a single library prep. Together with the streamlined workflow and possibility of requesting up to 384 barcodes, this unique kit allows the interrogation of both bacterial and micro-eukaryotic populations within a complex community taking advantage of the high sensitivity of NGS.

 

Optimized Protocol Offers Low PCR Bias, Fewer Off-target Reads, and Limited Cross-reactivity with Human Samples

The protocol calls for two steps, an initial PCR amplification of the specified target regions using customized PCR primers that target both the 16S V4 and 18S domains, and then a subsequent PCR amplification that adds a sample-specific index. Compared to more traditional sequencing methods, the incorporation of the second PCR step reduces off-target reads typically encountered during amplicon sequencing.

 

Automated Solution for 16S V4 & 18S rRNA Sequencing

The NEXTFLEX® 16S V4 & 18S Automation kit is a ready-to-go solution for automation on the PerkinElmer® Sciclone G3 NGS/NGSx workstation. The kit includes the library prep reagents with additional overage considerations and single-use, plug-and-play 96-barcode plates to enable a smooth transition for labs wanting a solution for their high-throughput needs. 

 

产品特点： 

• Multiplex up to 384 samples

• Fast and easy joint library prep protocol for both target regions

• Low input – Only 1 ng of input required

• Custom sequencing primers not required

• Automation protocols are available for the PerkinElmer Sciclone® NGS and NGSx Workstations

• Functionally validated on the Illumina® sequencing platforms

 

产品列表：

货号	产品名称	规格
NOVA-4213-01	NEXTFLEX® 16S V4 and 18S  Amplicon-Seq Kit-8 BARCODES	8 RXNS
NOVA-4213-02	NEXTFLEX® 16S V4 and 18S  Amplicon-Seq Kit-48 BARCODES	48 RXNS
NOVA-4213-13	NEXTFLEX® 16S V4 and 18S  Amplicon-Seq Kit -BARCODES 1-96	96 RXNS

 

KIT CONTENTS

• NEXTFLEX® PCR Master Mix
• NEXTFLEX® 16S V4 & 18S PCR I Primer Mix
• NEXTFLEX® PCR II Barcoded Primer Mix
• Resuspension Buffer
• Nuclease-free Water
• NEXTFLEX® Cleanup Beads

REQUIRED  MATERIALS NOT PROVIDED

• 1 ng – 50 ng high-quality environmental DNA in up to 34 µL nuclease-free water for each library
• 96 well PCR Plate Non-skirted (Phenix Research, Cat # MPS-499) or similar
• Adhesive PCR Plate Seal (Bio-Rad®, Cat # MSB1001)
• Magnetic Stand – 96 (Thermo Fisher® Scientific, Cat # AM10027) or similar
• Thermocycler
• 2, 10, 20, 200 and 1000 µL pipettes / multichannel pipettes
• Nuclease-free barrier pipette tips
• Vortex
• 80% Ethanol, freshly prepared (room temperature)

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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