Sequencing By Synthesis
Sequencing By Synthesis - Related Videos
09:34
Synthesis of Information-bearing Peptoids and their Sequence-directed Dynamic Covalent Self-assembly
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Cited by 2 •
2020
A protocol is presented for the synthesis of information-encoded peptoid oligomers and for the sequence-directed self-assembly of these peptoids into molecular ladders using amines and aldehydes as dynamic covalent reactant pairs and Lewis acidic rare-earth metal triflates as multi-role reagents.
02:36
Transfer RNA Synthesis
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2020
One of the unique features of tRNA is the presence of modified bases. In some tRNAs, modified bases account for nearly 20% of the total bases in the molecule. Altogether, these unusual bases protect the tRNA from enzymatic degradation by RNases. Each of these chemical modifications is carried by a specific enzyme, post-transcription. All of these enzymes have unique base and site-specificity. Methylation, the most common chemical modification, is carried by at least nine different enzymes, with...
09:42
Solid Phase Synthesis
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2023
Source: Vy M. Dong and Diane Le, Department of Chemistry, University of California, Irvine, CA Merrifield's solid-phase synthesis is a Nobel Prize winning invention where a reactant molecule is bound on a solid support and undergoes successive chemical reactions to form a desired compound. When the molecules are bound to a solid support, excess reagents and byproducts can be removed by washing away the impurities, while the target compound remains bound to the resin. Specifically, we will...
08:33
Hydrogel Synthesis
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2023
Source: Amber N. Barron, Ashlea Patterson, and Taylor D. Sparks, Department of Materials Science and Engineering, The University of Utah, Salt Lake City, UT Hydrogels are a versatile class of cross-linked polymers produced through relatively simple procedures and with generally inexpensive materials. They can be formed from solution and involve a polymer backbone formed from monomer reagents, an initiator which makes the polymer reactive and a crosslinking species which binds the polymer chains...
08:57
Amplicon Sequencing using the Long-Read Sequencing Technologies
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2025
This protocol was optimized for targeted deep sequencing of 18 drug-resistance regions in Mycobacterium tuberculosis using a long-read sequencing platform, followed by analysis with a tuberculosis-specific bioinformatics pipeline designed for long-read data.
FAQs
During each cycle, a complementary nucleotide is added to the growing DNA strand, and that incorporation produces a detectable fluorescent or chemical signal. The identity of the signal indicates which base was incorporated at that position. Repeating this process creates an ordered series of base calls, which is then converted into the DNA sequence.
Repeated cycles allow the method to determine the sequence one position at a time rather than measuring an entire DNA fragment at once. DNA fragments remain attached to a surface while successive complementary bases are incorporated. Recording the signal from each cycle preserves the order of the bases and supports parallel analysis of many fragments.
Surface attachment keeps DNA fragments positioned for repeated reactions and signal measurements. Because the fragments remain associated with the same analysis surface across cycles, signals can be tracked as new complementary nucleotides are incorporated. This arrangement supports parallel processing and contributes to the high-throughput capacity of the sequencing workflow.
A typical workflow begins by attaching DNA fragments to a surface. The system then performs repeated cycles of complementary nucleotide addition and detects the fluorescent or chemical signal associated with each incorporation. Finally, the ordered signal information is converted into DNA sequence data, producing results suitable for genome or transcript analysis.
Researchers apply this approach when they need sequence information from many DNA fragments, including genome analysis, transcript profiling, and variant detection. It can also support pathogen surveillance by revealing sequence patterns in biological samples. Its scalability makes it useful for projects requiring broad sequence coverage rather than analysis of only one fragment.
For variant detection, the resulting sequence data can be examined for differences within analyzed DNA relative to the expected sequence. In pathogen surveillance, sequence information helps characterize genetic material from pathogens and track relevant sequence patterns. These applications demonstrate how base-by-base measurements can support both biological interpretation and monitoring of infectious threats.