Next-generation sequencing (NGS), is a methodology that increases sequencing throughput by laying millions of DNA fragments on a single chip and sequencing all fragments in parallel. DNA fragments, which can be collected from various upstream processes, are used to build libraries that are then used as sequencing templates. These are prepared for sequencing by ligating specific adaptor oligonucleotides to both ends of each fragment. Following sequencing, informatics allows each sequencing read to be mapped to a reference genome.
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AIR™ Small RNA Sequencing Kit(Illumina Compatible)
Features The AIR™ Small RNA Sequencing Kit is designed to prepare small RNA libraries for sequencing usingIllumina®/Solexa® sequencers. Bioo Scientific’s AIR Small RNA Sequencing Kit streamlines the sample prep procedure required for the sequencing of microRNAs, piwi RNAs and other non-coding RNAs. The kit describes the methods required to prepare sample libraries to load onto flow cells for cluster generation.
Description This kit can be used to generate 10 small RNA libraries. There are four steps involved in preparing small RNAs for sequencing: RNA extraction and enrichment, adapter ligation, first strand synthesis, and PCR amplification. The kit contains the necessary material to take the user’s purified RNA sample through preparation and amplification for loading onto flow cells for sequencing. Download Manual
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AIR™ Ligase(T4 RNA Ligase 2, truncated)
Description Bioo Scientific’s AIR™ Ligase was developed specifically for demanding Next-Generation RNA Sequencing applications. AIR Ligase specifically ligates the adenylated 5´ end of an adapter to the 3´ end of RNA. The enzyme does not require ATP for ligation but does need an adenylated substrate, which dramatically reduces the amount of ligation between random RNA molecules. AIR Ligase is atruncated version of T4 RNA Ligase 2. Unlike the full length ligase, AIR Ligase does not ligate the phosphorylated 5´ end of RNA or DNA without the adenylated substrate making it an excellent choice for small RNA library preparation.Whether you plan to sequence microRNAs or perform directional mRNA-Seq, AIR Ligase will enhance your library preparations. This enzyme is compatible with all of our AIR™ Sequencing Grade products. For use withIllumina, SOLiD (ABI), 454 FLX (Roche) and any other NGS platform.White paper Get a free sample!
Benefits • Truncation makes downstream assays more robust
• Efficiency of ligation nears 100%
• Increase ability to identify miRNAs
AIR™ Ligase is much more efficient than the competitor’s T4 RNA Ligase 2, truncated.AIR™ Ligase ligation of an 25-mer microRNA with a 24-mer AIR™ Adenylated Linker. Lane 1 demonstrates a ligation using AIR™ Ligase (200 U) with supplied buffer. Lane 2 (200U) and lane 3 (400U) are examples of a competitor’s T4 RNA Ligase 2, truncated with supplied buffer.
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AIR™ Barcoded Adapters for Sample Multiplexing in Next Generation Sequencing (NGS)
Features AIR™ Barcoded Adapters significantly increase scale and throughputin small RNA NGS reactions, while reducing costs by allowing the user to pool multiple library preparations into a single sequencing reaction (sample multiplexing). AIR™ Barcoded Adapters can be used to multiplex NGS reactions for miRNA, piwi RNA (piRNA), small nucleolar RNA (snoRNA) and other small, noncoding RNAs, as well as for genomic DNA Sequencing Oligo, multiplexing DNA Sequencing Oligo, paired-End Sequencing Oligo and DpnII and NlAIII Gene Expression. REFERENCES: Lau, et al. Science 2001 294: 858-62.
“Sample barcoding technology maximizes Next-Generation Sequencing by allowing sample multiplexing, large library generation, cross-genomic and gene-drug treatment studies. This tagged technology improves sequencing experimental design to such a magnitude; it is likely all future Next-Generation Sequencing runs will be performed with barcodes.”
Dr. Masoud Toloue, Senior Scientist at Bioo Scientific
Description AIR™ Barcoded Adapters are 5' pre-adenylated 3’ adapters that are blocked at their 3’ends. This eliminates the need for ATP during ligation, minimizing ligation to the 5’ phosphate of the small RNA pool and subsequent circularization. Enough predesigned AIR™ Barcoded Adapters is provided for ten sequencing reacions per adapter.
Pre-designed AIR™ Barcoded Adapters:are available in sets of 10, 20, or 30 barcodes and are designed for multiplexing small RNA libraries for sequencing using Illumina®/Solexa® sequencers. Pre-designed AIR™ Barcoded Adapters were designed based on uniform melting temperature (Tm)and sequences that have unique color space. Once attached to a sample, the user may pool up to 10 samples after the 3’ adapter and 5’ adapter ligations. Alternatively, the user can pool up to 10 samples post PCR amplification. The kit contains the necessary barcoded adapters and primers for 25 reactions per barcode. Analysis post run allows the user to identify the sequence barcode back to specific samples. Download Manual
Custom AIR™ Barcoded Adapters: Bioo Scientific also offers custom barcoded adapters which can be designed to meet any of your specifications. Like the predesigned AIR Barcoded Adapters, the custom AIR Barcoded adapters are 5’adenylated barcoded linkers that are modified to prevent self-ligation and to improve efficiency. Custom adaptors can be designed for use with Illumina, SOLiD (ABI), 454 FLX (Roche) or any other next-generation sequencing platform. Phosphorothioate, adenylation, blocking groups and other modifications are available.
Benefits • allows sample multiplexing in small RNA NGS reactions: pool multiple libraries into a single reaction!
• allows to perform NGS with multiple samples, conditions and time-courses and even across multiple genomes
• increases sequencing scale and throughput and maximizes your flow cell
• reduces your per-sample cost by bar-coded multiplexing (affordable alternative to adapters found in sequencing kits!)
• Phosphorothioate, adenylation, blocking groups and other modifications available
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AIR™ Adenylated Linkers
Description AIR™ Adenylated Linkers are activated 5’adenylated oligos with 3’end blocks, that facilitate NGS and miRNA cloning and that preclude the need for ATP in RNA Ligase 2 (RNL2) reactions. This allows for specific and clean ligations. The 3’ terminal dideoxy C (ddC) modification prevents self-ligation and improves efficiency. Both the custom and pre-designed AIR™ Adenylated Linkers are highly cost effective. REFERENCES: Lau, et al. Science 2001 294: 858-62.
Predesigned AIR™ Adenylated Linkers:are predesigned 5’adenylated oligos with 3’end blocks, which are ready for use in next generation sequencing or miRNA cloning. Compatible withIllumina and 454 FLX (Roche).
AIR™ Adenylated Linker Acontains theBan-1 restriction site sequence
5’rAppCTGTAGGCACCATCAAT/3ddC/3’
AIR™ Adenylated Linker Bcontains the Ava-1 and Sty-1 restriction sequence sites
5’rAppCACTCGGGCACCAAGGA/3ddC/3’
AIR™ Adenylated Linker Ccontains the EcoR-1 restriction site sequence
5’rAppTTTAACCGCGAATTCCAG/3ddC/3’
Custom AIR™ Adenylated Linkers:are customized 5’adenylated oligos with 3’end blocks, that facilitate NGS and miRNA cloning. Design your own miRNA cloning linkers! Can be designed for use with Illumina, SOLiD (ABI), 454 FLX (Roche) or any other NGS platform. Phosphorothioate and other modifications available. Most custom AIR Adenylated Linkers have a 10 day turnaround time.
Benefits • Ideal for next-generation sequencing, miRNA cloning and miRNA sequencing
• 3’ end blocks prevent self-ligation
• Adenylated 5' end reacts with the truncated T4 RNL2 Ligase, no need for ATP!
• Reduces cost of NGS: affordable alternative to other adapter suppliers
• Phosphorothioate and other modifications available
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miR-Mouse™ Enriched Small RNAs
Description miR-Mouse™ Enriched Small RNAs:purified from pools of up to 10 organs from Balb mice at the age of either 8-10 weeks or 4-5 months. The RNA is extracted using a single-phase reagent containing phenol and guanidinium thiocyanate and the small RNA fraction is enriched by differential precipitation with isopropanol. The large RNA-enriched fraction corresponding to the particular small RNA-enriched preparation is available as a companion product. The small RNA enriched RNA ranges in size from approximately 10 bp to 100 bp and is especially useful for analyzing microRNA and other endogenous small RNAs in particular tissues. The product is supplied at a concentration of 50 ng/uL in a buffer of 0.2 mM EDTA pH 8 in nuclease-free water.
Custom miR-Mouse™ Enriched Small RNAs:The RNA is also available as a custom product of RNA from strain/gender/age as specified by the researcher.
Benefits • High-quality enriched small RNA
• Eliminates tedious, time-consuming lab procedures
• can be used fo Gene expression analysis, Microarray analysis, miRNA cloning
• Compatible withIllumina, SOLiD (ABI), 454 FLX (Roche) and any other NGS platforms
