Next® Single Cell/Low Input cDNA Synthesis & Amplification Module |NEB酶试剂 New England Biolabs

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产品信息

The NEBNext® Single Cell/Low Input cDNA Synthesis & Amplification Module uses a template switching method to generate full length cDNAs directly from single cells or 2 pg – 200 ng RNA. This unique workflow enables generation of high quality sequencing data from a broad range of inputs, and superior transcript detection, while providing reliably consistent performance.

Features:

  • The highest yields from single cells, or 2 pg – 200 ng RNA
  • Input can be cultured or primary cells, or total RNA
  • Single-tube protocol from cell lysis to cDNA
  • Fast, streamlined, automation-friendly workflow, with minimal handling steps and hands-on time

Download extensive performance data in our technical note.

Figure 1: NEBNext Single Cell/Low Input workflow


Next® Single Cell/Low Input cDNA Synthesis & Amplification Module |

Figure 2: Higher cDNA yields with the NEBNext Single Cell/ Low Input cDNA Synthesis & Amplification Module


Next® Single Cell/Low Input cDNA Synthesis & Amplification Module |

Sequencing libraries were generated from HeLa, Jurkat and M1 single cells, or 10 pg of Universal Human Reference (UHR) RNA (Agilent® #740000) with recommended amounts of ERCC RNA Spike-In Mix I (Thermo Fisher Scientific® #4456740). The NEBNext Single Cell/Low Input cDNA Synthesis & Amplification Module or the SMART-Seq® v4 Ultra® Low Input RNA Kit for Sequencing (Clontech® # 634891) were used. cDNA libraries were amplified using the number of PCR cycles shown. Error bars indicate standard deviation for 8-21 replicates.

 

View extensive performance data in our technical note.

产品类别:
RNA Library Prep for Illumina

  • 产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • E6421S     -20    
        NEBNext® Single Cell RT Primer Mix E6422AVIAL -20 1 x 0.048 ml Not Applicable
        NEBNext® Single Cell RT Buffer E6423AVIAL -20 1 x 0.12 ml Not Applicable
        NEBNext® Template Switching Oligo E6424AVIAL -20 1 x 0.024 ml Not Applicable
        NEBNext® Single Cell RT Enzyme Mix E6425AVIAL -20 1 x 0.048 ml Not Applicable
        NEBNext® Single Cell cDNA PCR Master Mix E6426AVIAL -20 1 x 1.2 ml Not Applicable
        NEBNext® Single Cell cDNA PCR Primer E6427AVIAL -20 1 x 0.048 ml Not Applicable
        NEBNext® Cell Lysis Buffer E6428AVIAL -20 1 x 0.048 ml Not Applicable
        Murine RNase Inhibitor E6429AVIAL -20 1 x 0.024 ml Not Applicable
        NEBNext® Bead Reconstitution Buffer E6430AVIAL -20 1 x 1.08 ml Not Applicable
        TE Buffer E6432AVIAL -20 1 x 1.2 ml Not Applicable
        Nuclease-free Water E6433AVIAL -20 1 x 1.44 ml Not Applicable
    • E6421L     -20    
        NEBNext® Single Cell RT Primer Mix E6422AAVIAL -20 1 x 0.192 ml Not Applicable
        NEBNext® Single Cell RT Buffer E6423AAVIAL -20 1 x 0.48 ml Not Applicable
        NEBNext® Template Switching Oligo E6424AAVIAL -20 1 x 0.096 ml Not Applicable
        NEBNext® Single Cell RT Enzyme Mix E6425AAVIAL -20 1 x 0.192 ml Not Applicable
        NEBNext® Single Cell cDNA PCR Master Mix E6426AAVIAL -20 1 x 4.8 ml Not Applicable
        NEBNext® Single Cell cDNA PCR Primer E6427AAVIAL -20 1 x 0.192 ml Not Applicable
        NEBNext® Cell Lysis Buffer E6428AAVIAL -20 1 x 0.192 ml Not Applicable
        Murine RNase Inhibitor E6429AAVIAL -20 1 x 0.096 ml Not Applicable
        NEBNext® Bead Reconstitution Buffer E6430AAVIAL -20 1 x 4.32 ml Not Applicable
        TE Buffer E6432AAVIAL -20 1 x 4.8 ml Not Applicable
        Nuclease-free Water E6433AAVIAL -20 1 x 5.76 ml Not Applicable

  • 特性和用法

    需要但不提供的材料

    80% Ethanol (freshly prepared)
    Nuclease-free Water
    DNA LoBind Tubes (Eppendorf® #022431021)
    Magnetic rack or plate (e.g., NEBNext® Magnetic Separation Rack (NEB #S1515S), Alpaqua ® 96S Super Magnet Plate (#A001322), or equivalent)
    Thermocycler
    Vortex
    Microcentrifuge
    SPRIselect Reagent Kit (Beckman Coulter®, Inc. #B23317) or AMPure® XP
    Beads (Beckman Coulter, Inc. #A63881)
    Agilent® Bioanalyzer® or similar fragment analyzer and associated consumables
    DNase RNase free PCR strip tubes (USA®  Scientific 1402-1708)

操作说明、说明书 & 用法

  • 说明书

    产品说明书包含产品使用的详细信息、产品配方和质控分析。

    • manualE6421

FAQs & 问题解决指南

  • FAQs

    1. How much PBS can be carried over from cell collection into the cDNA Synthesis and Amplification protocol?
    2. What cell types does the NEBNext Single Cell Low Input cDNA Synthesis Amplification Module support
    3. Can I resuspend the cells in buffers medium other than PBS before cell collection
    4. Do the cells have to be sorted in to a specific reagent
    5. Can frozen cells be used?
    6. Can cells be lysed using other cell lysis buffers
    7. How long can the synthesized cDNA (prior to cDNA amplification) be stored for prior to amplification
    8. What types of RNA are compatible
    9. Does the input total RNA have to be purified
    10. My total RNA sample is in a volume 8 µl Can I still use the module
    11. Is the cDNA generated by the NEBNext Single Cell Low Input cDNA Synthesis Amplification protocol compatible with Nextera XT from Illumina
    12. How should the adaptor sequences be trimmed
    13. Where can I find protocols for low-input RNA library prep for use with the following kits: NEBNext® Single Cell/Low Input RNA Library Prep Kit for Illumina (NEB #E6420), NEBNext® Single Cell/Low Input cDNA Synthesis & Amplification Module (NEB #E6421), and NEBNext® Single Cell Lysis Module (#E5530)?