Next® Globin & rRNA Depletion Kit (Human/Mouse/Rat) with RNA Sample Purification Beads |NEB酶试剂 New England Biolabs

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产品信息

The great majority of RNA in blood samples is comprised of cytoplasmic and mitochondrial ribosomal RNAs (rRNA) and globin mRNA. These highly abundant RNA species can conceal the biological significance of less abundant transcripts, and so their efficient and specific removal is desirable.

The NEBNext® Globin & rRNA Depletion Kit (Human/Mouse/Rat) employs the NEBNext RNase H-based RNA depletion workflow to deplete adult, embryonic and fetal globin mRNA (HBA1/2, HBB, HBD, HBM, HBG1/2, HBE1, HBQ1 and HBZ), cytoplasmic rRNA (5S, 5.8S, 18S, 28S, ITS and ETS) and mitochondrial rRNA (12S and 16S).

The kit is effective with human, mouse and rat total RNA preparations, both intact and degraded. The resulting depleted RNA is suitable for RNA-seq, random-primed cDNA synthesis, or other downstream RNA analysis.

This kit can also be used following poly(A) mRNA enrichment (e.g., using the NEBNext poly(A) mRNA Magnetic Isolation Module NEB #E7490), so that the final depleted RNA contains only mRNA of interest and no non-coding RNA. 
 
The kit is also available without RNAClean® beads.
 
Features

  • Efficient, specific depletion of rRNA and globin mRNA (adult, fetal and embryonic)
  • Suitable for low-quality or high-quality RNA
  • Compatible with a broad range of input amounts: 10 ng – 1 μg
  • Optional integration with poly(A) mRNA isolation workflows for removal of globin RNAs, rRNAs, and noncoding RNAs
  • Fast workflow: 2 hours, with less than 10 minutes hands-on time

 

 

Figure 1. Depletion of globin mRNA and ribosomal RNA enriches for RNAs of interest across species

Next® Globin & rRNA Depletion Kit (Human/Mouse/Rat) with RNA Sample Purification Beads |

Human, mouse and rat whole blood total RNA (1 µg) was depleted of rRNA alone, or rRNA and globin mRNA transcripts, using the NEBNext Globin & rRNA Depletion Kit.  RNA-seq libraries were prepared from untreated and depleted RNA using the NEBNext Ultra™ II RNA Library Prep Kit for Illumina® followed by paired-end sequencing (2 x 75 bp). Reads were identified as rRNA or globin mRNA using mirabait (6 or more, 25-mers), and levels of rRNA and globin mRNA remaining were calculated by dividing matched reads by the total number of reads passing instrument quality filtering.
Figure 2: Consistent Depletion Across Species and Across Inputs

Next® Globin & rRNA Depletion Kit (Human/Mouse/Rat) with RNA Sample Purification Beads |

Human, mouse and rat whole blood total RNA (1 µg, 100 ng and 10 ng) was depleted of rRNA and globin mRNA using the NEBNext Globin & rRNA Depletion Kit or Globin-Zero® Gold rRNA Depletion Kit (Illumina®). RNA-seq libraries were prepared from untreated and depleted RNA using the NEBNext Ultra™ II RNA Library Prep Kit for Illumina® followed by paired-end sequencing (2 x 75 bp). Reads were identified as ribosomal or globin using mirabait (6 or more, 25-mers), and levels of rRNA and globin mRNA remaining were calculated by dividing matched reads by the total number of reads passing instrument quality filtering. The data represents an average of 3 replicates and error bars indicate standard error. The NEBNext Globin Depletion Kit is superior at depleting rRNA across species, and at depleting over 99% of globin mRNA.
Figure 3: Transcript expression correlation is maintained after depletion

Next® Globin & rRNA Depletion Kit (Human/Mouse/Rat) with RNA Sample Purification Beads |

Human whole blood total RNA (1 µg) was depleted of rRNA and globin mRNA using the NEBNext Globin & rRNA Depletion Kit or Globin-Zero® Gold rRNA Depletion Kit (Illumina®). RNA-seq libraries were prepared from untreated and depleted RNA using the NEBNext Ultra™ II RNA Library Prep Kit for Illumina® followed by paired-end sequencing (2 x 75 bp). GENCODE v27 transcript abundances were estimated using Salmon. TPM (Transcript Per Million mapped reads) of protein coding transcripts, and R2 values for the linear fit are shown. Correlation analysis of the transcripts indicates better transcript expression correlation between depleted and undepleted samples for the NEBNext Globin & rRNA Depletion Kit. Treatment does not alter the abundances of non-targeted transcripts.
Figure 4: Transcript expression correlation is maintained across a range of input amounts

Next® Globin & rRNA Depletion Kit (Human/Mouse/Rat) with RNA Sample Purification Beads |

Human whole blood total RNA (1 µg, 100 ng and 10 ng) was depleted of rRNA and globin mRNA using the NEBNext Globin & rRNA Depletion Kit. RNA-seq libraries were prepared using the NEBNext Ultra™ II RNA Library Prep Kit for Illumina® followed by paired-end sequencing (2 x 75 bp). GENCODE v27 transcript abundances were estimated using Salmon. TPM (Transcript Per Million mapped reads) and R2 values for the linear fit are shown. Correlation analysis of the transcripts indicates consistent transcript expression regardless of input amount.
产品类别:
RNA Depletion & mRNA Enrichment Products,
RNA Library Prep for Illumina,
Next Generation Sequencing Library Preparation Products

  • 试剂盒组成

    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
    • E7755S     Multi-temperature    
        NEBNext® Globin & rRNA Depletion Kit (Human/Mouse/Rat) E7750S -20    
        NEBNext® DNase I E7753-2VIAL -20 1 x 0.015 ml 20,000 units/ml
        NEBNext® Thermostable RNase H E7752-2VIAL -20 1 x 0.012 ml 5,000 units/ml
        NEBNext® Globin and rRNA Depletion Solution E7751-2VIAL -20 1 x 0.018 ml 1 X
        RNase H Reaction Buffer E6312-2VIAL -20 1 x 0.012 ml Not Applicable
        DNase I Reaction Buffer E6315-2VIAL -20 1 x 0.03 ml Not Applicable
        Nuclease-free Water E6317-2VIAL -20 1 x 0.4 ml Not Applicable
        NEBNext Probe Hybridization Buffer E6314-2VIAL -20 1 x 0.012 ml Not Applicable
        NEBNext® RNA Sample Purification Beads E6351S 4    
        NEBNext® RNA Sample Purification Beads E6351SVIAL 4 1 x 0.66 ml Not Applicable
    • E7755L     Multi-temperature    
        NEBNext® Globin & rRNA Depletion Kit (Human/Mouse/Rat) E7750L -20    
        NEBNext® DNase I E7753-3VIAL -20 1 x 0.06 ml 20,000 units/ml
        NEBNext® Thermostable RNase H E7752-3VIAL -20 1 x 0.048 ml 5,000 units/ml
        NEBNext® Globin and rRNA Depletion Solution E7751-3VIAL -20 1 x 0.072 ml 1 X
        RNase H Reaction Buffer E6312-3VIAL -20 1 x 0.048 ml Not Applicable
        DNase I Reaction Buffer E6315-3VIAL -20 1 x 0.12 ml Not Applicable
        Nuclease-free Water E6317-3VIAL -20 1 x 1.5 ml Not Applicable
        NEBNext Probe Hybridization Buffer E6314-3VIAL -20 1 x 0.048 ml Not Applicable
        NEBNext® RNA Sample Purification Beads E6351L 4    
        NEBNext® RNA Sample Purification Beads E6351LVIAL 4 1 x 2.64 ml Not Applicable
    • E7755X     Multi-temperature    
        NEBNext® Globin & rRNA Depletion Kit (Human/Mouse/Rat) E7750X -20    
        NEBNext® DNase I E7753-4VIAL -20 1 x 0.24 ml 20,000 units/ml
        NEBNext® Thermostable RNase H E7752-4VIAL -20 1 x 0.192 ml 5,000 units/ml
        NEBNext® Globin and rRNA Depletion Solution E7751-4VIAL -20 1 x 0.288 ml 1 X
        RNase H Reaction Buffer E6312-4VIAL -20 1 x 0.192 ml Not Applicable
        DNase I Reaction Buffer E6315-4VIAL -20 1 x 0.48 ml Not Applicable
        Nuclease-free Water E6317-4VIAL -20 1 x 6 ml Not Applicable
        NEBNext Probe Hybridization Buffer E6314-4VIAL -20 1 x 0.192 ml Not Applicable
        NEBNext® RNA Sample Purification Beads E6351X 4    
        NEBNext® RNA Sample Purification Beads E6351XVIAL 4 1 x 10.6 ml Not Applicable

  • 特性和用法

    需要但不提供的材料

    • Magnetic rack/stand such as the NEBNext Magnetic Separation Rack (NEB #S1515)
    • 80% Ethanol (freshly prepared)
    • Thermocycler
    • Thin wall 200 µl PCR tubes
    • Bioanalyzer®, TapeStation® (Agilent Technologies, Inc.) or similar fragment analyzer and consumables

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。

    • manualE7750_E7755

工具 & 资源

  • Web 工具

    • NEBNext Selector

FAQs & 问题解决指南

  • FAQs

    1. What species can I use the NEBNext® Globin mRNA & rRNA Depletion Kit for? Will it work for other species?
    2. Which globin mRNAs and rRNAs are depleted with the NEBNext® Globin mRNA & rRNA Depletion Kit (Human/Mouse/Rat)? 
    3. Can I use the NEBNext® Globin mRNA & rRNA Depletion Kit (Human/Mouse/Rat) with degraded RNA or fragmented RNA?
    4. What is the total RNA input I should use?
    5. Why must the RNA be free of DNA?
    6. What is the expected RNA yield recovered after globin mRNA and rRNA depletion?
    7. What is the percentage of globin mRNA and rRNA left after depletion?
    8. How can I determine if the globin mRNA and rRNA depletion was efficient?
    9. I am only interested in coding RNA. Can I use the NEBNext® Globin mRNA & rRNA Depletion Kit (Human/Mouse/Rat) to remove globin mRNA from previously enriched mRNA.
    10. Does the NEBNext® Globin mRNA & rRNA Depletion Kit (Human/Mouse/Rat) work for ribosomal footprinting or profiling applications?
    11. Can the enriched RNA resulting from the NEBNext depletion kit be used in long read sequencing applications?