Small RNAs act through various pathways, including mRNA degradation, translational repression, heterochromatin formation, and DNA removal to regulate organismal growth and development. Lifeasible can help you obtain genome-wide small RNA profiles of species by constructing small RNA libraries and performing large-scale sequencing of small RNA libraries. This includes the mining of new small RNA molecules, prediction and identification of target genes, differential expression analysis between samples, small RNA clustering, and expression profiling.

With the development of second-generation sequencing technology, small RNA libraries constructed using different library construction techniques are suitable for different sequencing platforms, and complete genomics (CG) sequencing is one of them. It uses high-density DNA nano-chip technology, in which DNA nanospheres are embedded in the chip, and the sequence is read with cPAL technology.

We provide library construction methods for small RNA that can be used for library construction of small RNA from all sources, and the constructed small RNA CG libraries are suitable for CG sequencing platforms, which offer higher throughput and low cost per genome sequenced. The CG library construction-based sequencing technology we provide is characterized by low library construction cost, simple operation, short time, a high success rate of library construction, and fast sequencing.

Construction of a small RNA CG library

• Small RNA fragments are connected to the 3' junction of the CG library.
• A small RNA fragment connects to the 5' junction of the CG library.
• The small RNA attached to the 3' and 5' junctions of the CG library is reverse transcribed into cDNA.
• PCR amplification of the reverse transcription product.
• Purify and recover the PCR amplified product by non-denaturing PAGE gel (e.g., 6% PAGE gel).
• Qubit quantitative purification of the recovered PCR products.
• Single-strand separation of purified and recovered PCR products.
• Cyclization of single-stranded DNA.
• Enzymatic digestion of uncyclised single-stranded DNA.
• Purification and recovery of digested products (e.g., using PEG32 beads).
• Library quality control and concentration determination.

Sequencing of small RNA CG library and downstream data analysis

• Small RNA CG library CG sequencing platform on-line sequencing.
• Analysis of small RNA CG library downstream data.

Differences and advantages between our solution and DNA CG library construction

• DNA CG library construction requires dephosphorylation and end-flattening of the constructed fragments, which is not required to construct our small RNA CG libraries.
• The 3' and 5' junctions in the DNA CG library construction scheme are attached to both ends of the library construct in the same reaction system. In contrast, the 3' and 5' junctions of our small RNA library construction are attached to both ends of the library fragment in two separate reaction systems, and the 3' and 5' junctions are designed and synthesized independently by us for small RNA.
• The 3' and 5' junctions are designed and synthesized by us for small RNA. The construction of DNA CG libraries requires an immediate nick-panning reaction after attachment of the junctions and then PCR amplification. In contrast, the construction of small RNA CG libraries we provided does not require the nick-shift reaction. It only requires reverse transcription of the library fragments attached to the junction before PCR amplification.
• The construction of the DNA CG library can be processed by single-strand separation after PCR amplification. For the construction of small RNA CG libraries, it is necessary to use non-denaturing PAGE gel to purify and recover the PCR amplification products before single-strand separation.

Through extensive and in-depth research and extensive screening, Lifeasible provides customers with vesicular junctions for the efficient preparation of high-quality nucleic acid sequencing libraries that can be used in nucleic acid sequencing library construction. The experimental results have demonstrated that the library quality and relevance of the sequencing library constructed from the vesicular junction is very high compared to the sequencing data obtained from other nucleic acid sequencing library construction techniques and can be used in CG sequencing platforms. The data obtained is of high authenticity and credibility and has no impact on the analysis of the information. Please feel free to contact us for more information.

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For research or industrial use.

Related Services

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