The ABclonal mRNA-seq Lib Prep Kit for Illumina ® provides a method for generating non-directional (non-strand-specific) RNA library , which contains Poly(A) mRNA Purification Module(ABclonal, Cat. RK20341), mRNA-seq Lib Prep Module for Illumina®( ABclonal, Cat. RK20350) and RNA adaptors for Illumina. The kit is applicable to animal, plant, and eukaryote (e.g., fungi) total RNA templates as well as the isolated and purified mRNA templates. The recommended total RNA input is 10 ng-1 μg.
Workflow Chart
Storage
Box-1: 2-8°C; Box-2: -20°C; Box-3: -20°C.
Product Information
Product Components
Box
Box Name
Tube Name
24 rxns
96 rxns
Box 1
Poly(A) mRNA Purification Module (RK20341)
Oligo d(T)25 Capture Beads
480 μl
1920 μl
mRNA Binding Buffer
12 ml
48 ml
Washing Buffer
19.2 ml
76.8 ml
Tris Buffer
1.2 ml
4.8 ml
Box 2
mRNA-seq Lib Prep Module for Illumina (RK20350)
2X Frag/Elute Buffer
144 μl
576 μl
RT Reagent
192 μl
768 μl
First Strand Synthesis Enzyme Mix
48 μl
192 μl
Second Strand Synthesis Reaction Buffer
192 μl
768 μl
Second Strand Synthesis Enzyme Mix
96 μl
384 μl
Nuclease-free Water
1 ml X 2
8 ml
End Prep Buffer
240 μl
960 μl
End Prep Enzyme Mix
72 μl
288 μl
Ligation Buffer
396 μl
1584 μl
Ligase Mix
72 μl
288 μl
2X PCR Mix
600 μl
2×1200 μl
Low-EDTA TE
1 mL X 3
10 ml
Box 3
RNA Adapter Module (24 Indices)(RK20345)
RNA Truncated Adapter
60 μl
240 μl
RNA Universal Primer
60 μl
240 μl
RNA Index Primer
2.5 μl*
10 μl*
Product Data
1.ABclonal mRNA-seq libraries showed high efficiency of first cDNA synthesis and library yield
10 ng, 100ng or 1 μg of Total RNA was used as input for libraries after using Poly(A) mRNA Purification Module (ABclonal, Cat. RK20341), and libraries were then prepared using mRNA-seq Lib Prep kits from different manufacturers. ABclonal mRNA-seq Lib Prep Module for Illumina showed significantly higher efficiency of first cDNA synthesis (left panel) and library yield (right panel) than competitors.
2.ABclonal mRNA-seq libraries retain highly correlation of transcript expression between different total RNA inputs
Poly(A)-containing mRNA was isolated from 10 ng, 100ng or 1 μg of Total RNA, and libraries were prepared using the ABclonal mRNA-seq Lib Prep Kit. Libraries were sequenced on an Illumina platform. FPKM correlation analysis of the transcripts indicates excellent correlation of transcript expression between different total RNA inputs.
3.ABclonal mRNA-seq libraries retain high ECRR transcript expression correlation
10 ng, 100ng or 1 μg of Total RNA was mixed into the same amount of ERCC RNA Spike-in Control Mixes(ambion, 4456740) and libraries were then prepared using the mRNA-seq Lib Prep kits from different manufacturers. Libraries were sequenced on an Illumina platform. ABclonal mRNA-seq Lib Prep Kit showed significantly higher ERCC transcript correlation than competitors.
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