Suitable for Illumina sequencing platforms;
The rRNA depletion module (H/M/R) is mainly for human, mouse and rat total RNA, which can effectively remove cytoplasmic rRNA from total RNA (including cytoplasmic 5S rRNA, 5.8S rRNA, 18S rRNA and 28S rRNA) and mitochondrial rRNA (containing 12S rRNA and 16S rRNA).
This kit is not only suit for complete RNA samples with good integrity, but also for RNA samples that have been degraded, such as FFPE RNA. The total amount of input RNA is 10 ng to 1 ug.
Stranded mRNA Lib Prep method is recommended as a follow strategy, Whole RNA-seq data can be used to analyze all mRNAs except ribosomes as well as some long-chain non-coding RNAs, stranded Library can quantify gene expression and RNA species more accurately, and more fully understand gene structure.
The kit contains an RNA truncated adapter, uses RNA index primer for PCR to add a unique index to each sample. Truncated adapter has higher connection efficiency than full length adapter and can reduce the generation of Adapter dimer;
ABclonal Whole RNA-seq Lib Prep kit for illumine contains rRNA depletion module (H/M/R), First Strand synthesis module（stranded），Second cDNA synthesis module（stranded）, DNA Lib Prep module with UDG and ABclonal RNA index primer. Reagents and enzymes for Whole RNA-seq Library preparation are all contained.
Each reagent in the kit has undergone rigorous quality control, and each lot of kits has been verified by Lib preparation and sequencing, ensuring stable quality performance of each lot of kits.
The following workflow illustrates the processes of the Rapid DNA Lib Prep Kit.
|Box||Module||Name and color||24 Reactions (RK20303M)||96 Reactions (RK20303L)|
|Box-1||rRNA Depletion Module (H/M/R)（RK20348）||Probe Hybridization Buffer||48 μL||192 μL|
|rRNA Probe Mix(H/M/R)||24 μL||96 μL|
|RNase H||48 μL||192 μL|
|10× RNase H Buffer||48 μL||192 μL|
|DNase I||60 μL||240 μL|
|10× DNase I Buffer||120 μL||480 μL|
|Box-2||First StrandSynthesis Module (Stranded) (RK20342)||2X Frag/Elute Buffer||264 μL||1056 μL|
|RT Strand Specificity Reagent||192 μl||768 μl|
|First Strand Synthesis Enzyme Mix||48 μl||192 μl|
|Second Strand Synthesis Module (Stranded) (RK20343)||Second Strand Synthesis Reaction Buffer with dUTP||192 μl||768 μl|
|Second Strand Synthesis Enzyme Mix||96 μl||384 μl|
|Nuclease-free water||2 ml||8 ml|
|DNA Lib Prep Module with UDG (RK20344)||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|
|UDG Enzyme||12 μl||48 μl|
|Low-EDTA TE||2.5 ml||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*|
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