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RNA Sequencing

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RNA sequencing (RNA-Seq) is a powerful technique that allows researchers to study the complete transcriptome profiling. A transcriptome is a complete set of transcripts, which includes their quantities as available in the cell, for specific physiological conditions. A complete understanding of the transcriptome gives enormous information on the functional components of the genome, molecular components of cell, tissues and knowledge on disease development.

Different technologies such as hybridization-based, and sequence-based approaches, and specialised microarrays have also been used earlier to study the transcriptome, but they were found to not be very effective for large volumes of data. Development of RNA-Seq based on the high-throughput DNA sequencing method has solved that problem to a great extent.

RNA Sequencing Technique

Every sample is precious, and data integrity is paramount. We communicate with our customers at every stage of the sequencing process, to ensure that they receive the maximum value out of every sample.

We also recognize that the value our customers get is not in the sequencing alone; it’s the technologies within the sequencing. MedGenome is one of the few genomics research companies that provides turnkey solutions to our customers, offering robust bioinformatics services alongside NGS sequencing.

Sample Requirements
DNA RNA
CELLS Minimum 1e+6 cells required. Cells should be shipped in dry ice Minimum 1e+6 cells required. Cells should be frozen and pelleted and shipped in dry ice
TISSUE 30 mg of fresh or frozen tissue or 20 mg of stabilized tissue. The tissue should be immersed in Liq N2 and shipped in dry ice or should be

immersed in RNAlater RNA Stabilization Reagent or All protect Tissue Reagent

30 mg of fresh or frozen tissue or 20 mg of stabilized tissue. The tissue should be immersed in Liq N2 and shipped in dry ice or should be

immersed in RNAlater RNA Stabilization Reagent or All protect Tissue Reagent

FFPE 5 slides with 10um thick tissues and surface area of 250 mm2 8 slides with 10um thick tissues and surface area of 250 mm2
BLOCK Sufficient thickness to generate the optimal number of slides as required in FFPE row
whole genome sequencing 100 ng
whole exome sequencing 100 ng
RNA sequencing 0.2 ug of high quality RNA (RIN>8) or 1ug RNA (RIN<8)
RNA-TruSeq access libraries / Libraries from ffpe rna 10 ng of high quality RNA (RIN>8) or 50ng RNA (RIN<5)
truseq standed mrna library 100 ng high quality RNA (RIN>8) or 1ug RNA with RIN between 5 and 8.
Genome in a Bottle validation results for WGS & WES
Sample name analysis type variant type accuracy precision analytical sensitivity analytical specificity
Asian son (NA24631) WGS SNP 99.9972% 99.9972% 98.3605% 99.9993%
Asian son (NA24631) WGS INDEL 99.9990% 97.2113% 95.9767% 99.9996%
Asian son (NA24631) WES SNP 99.9987% 99.4271% 98.9305% 99.9996%
Asian son (NA24631) WES INDEL 99.9996% 97.4525% 94.9458% 99.9999%
Utah female (NA12878) WGS SNP 99.9986% 99.5894% 99.2871% 99.9995%
Utah female (NA12878) WGS INDEL 99.9991% 97.6290% 96.7742% 99.9996%
Utah female (NA12878) WES SNP 99.9990% 99.3678% 99.3030% 99.9995%
Utah female (NA12878) WES INDEL 99.9995% 93.7846% 96.4536% 99.9997%