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How Lynx s MPSS Technology Works, Step by Step

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1. Create ds cDNA from expressed mRNA.

2. Chop up cDNA with DpnII (GATC recognition sequence), then select for the 3’ most fragment that extends to the poly (A) addition site.

3. Tag each cDNA at its poly (A) end with a 32-base nucleotide tag consisting of the bases A, T, G and comprising eight different four-letter nucleotide words, for a total of 16.7 million different tags. To be 99 percent confident that each cDNA will be tagged with a unique tag, only 1 percent of the tags are used at once.

4. Amplify the tagged cDNA fragments using PCR.

5. Mix in the tagged fragments with Lynx’s microbeads, each of which has 32-base antitags radiating out from it. The antitags, made out of the 16.7 million different 32-base pair permutations of C,A, and T, are designed to be complementary to the tags. Each bead has at least 100,000 identical antitags on it, which are synthesized on the bead base by base.

6. The tags hybridize to the antitags. All of the tags and antitags have the same melting temperature, but the nearest neighbor mismatch for each tag has a Tm several degrees different than the perfect match. So temperatrue control limits non-specific binding.

7. The DNA-laden beads are rolled into a flow cell to create a monolayer, and then into a sequencing instrument. The sequencing reagents are added to the surface of the beads, and flow through the flow cell while the beads stay still.

8. The DNA on the beads is sequenced four bases at a time for 17 bases toward the center, the minimum number of bases to determine a unique sequence.

9. These sequences are identified and counted. The level of expression for one gene is expressed as no. identical signatures/total no. transcripts analyzed.

The Scan

Researchers Develop Polygenic Risk Scores for Dozens of Disease-Related Exposures

With genetic data from two large population cohorts and summary statistics from prior genome-wide association studies, researchers came up with 27 exposure polygenic risk scores in the American Journal of Human Genetics.

US Survey Data Suggests Ancestry Testing Leads Way in Awareness, Use of Genetic Testing Awareness

Although roughly three-quarters of surveyed individuals in a Genetics in Medicine study reported awareness of genetic testing, use of such tests was lower and varied with income, ancestry, and disease history.

Coral Genome Leads to Alternative Amino Acid Pathway Found in Other Non-Model Animals

An alternative cysteine biosynthesis pathway unearthed in the Acropora loripes genome subsequently turned up in sequences from non-mammalian, -nematode, or -arthropod animals, researchers report in Science Advances.

Mosquitos Genetically Modified to Prevent Malaria Spread

A gene drive approach could be used to render mosquitos unable to spread malaria, researchers report in Science Advances.