chemistry

Template-switching polymerase chain reaction

Template-switching polymerase chain reaction is a chemistry topic covered in the lgStudy science library. This page brings together a partial reference excerpt, illustrations, worked examples, real-world applications and a short study plan, so you can understand Template-switching polymerase chain reaction rather than just read about it. In short: Template-switching polymerase chain reaction (TS-PCR) is a method of reverse transcription and polymerase chain reaction (PCR) amplification that relies on a natural PCR primer sequence at the polyadenylation site, also known as the poly(A) tail, and adds a second primer through the activity of murine leukemia virus reverse transcriptase. This permits reading full cDNA sequences and can deliver high yield from singl…

Key takeaways

Template-switching polymerase chain reaction belongs to chemistry; place it in that map before memorising details.
Learn the definition first, then one example that makes the definition concrete.
Connect Template-switching polymerase chain reaction to a quantity you can measure, compute or draw — that is where exam questions come from.
Reproduce the core statement of Template-switching polymerase chain reaction from memory before moving on to harder problems.

Reference excerpt

Template-switching polymerase chain reaction (TS-PCR) is a method of reverse transcription and polymerase chain reaction (PCR) amplification that relies on a natural PCR primer sequence at the polyadenylation site, also known as the poly(A) tail, and adds a second primer through the activity of murine leukemia virus reverse transcriptase. This permits reading full cDNA sequences and can deliver high yield from single sources, even single cells that contain 10 to 30 picograms of mRNA, with relatively low levels (3-5%) of contaminating rRNA sequence. This technique is often employed in whole transcriptome shotgun sequencing. It is marketed by Clontech as Switching Mechanism At the 5' end of RNA Template (SMART) as well as by Diagenode as Capture and Amplification by Tailing and Switching (CATS).

Drop-Seq By using syringe pumps to transmit a steady rate of isolated cells and uniquely oligonucleotide-barcoded beads, it is possible to isolate individual cells and beads together in droplets of lysis buffer, where the polyadenylation site binds to a primer containing a unique, identifying sequence. This primer also contains a common sequence upstream of the identifier, so that after it is extended by reverse transcription, subsequent rounds of PCR will incorporate the tag, which permits each isolated cDNA that is sequenced to be tracked back to a specific originating bead. This permits the relative levels of transcripts in many individual cells to be analyzed simultaneously, creating a rational basis for the classification of these cells into particular cell types, or permitting the logical inference of in situ hybridization data from embryos without actually performing the experiment.

References

Worked examples

Example 1 — a first encounter with Template-switching polymerase chain reaction

Start with the simplest possible case. Write down what Template-switching polymerase chain reaction claims or describes in one sentence, then invent the smallest concrete situation in which that sentence is true. In chemistry, the smallest case is usually a single object, a single equation or a single measurement. Check that every symbol or term in your sentence has a meaning in that case.

Example 2 — changing one variable

Take the situation from Example 1 and change exactly one quantity: double it, halve it, or set it to zero. Predict what should happen to Template-switching polymerase chain reaction before you calculate. Comparing your prediction with the result is the fastest way to find out whether you understand the idea or only the words.

Example 3 — an exam-style question

Typical questions about Template-switching polymerase chain reaction ask you to (a) state it precisely, (b) apply it to given data, and (c) explain a limitation. Practise writing all three answers in under five minutes; the third part is what separates a full-mark answer from an average one.

Applications of Template-switching polymerase chain reaction

In research: Template-switching polymerase chain reaction appears in chemistry research whenever the underlying quantities have to be modelled precisely. Papers usually cite it as a starting assumption and then explore where it breaks down.
In technology and industry: Engineering practice reuses Template-switching polymerase chain reaction in design rules, simulations and safety margins. Knowing the idea lets you read a specification sheet and understand why the numbers look the way they do.
In the classroom: Template-switching polymerase chain reaction is common in secondary-school and first-year university syllabi. It links to neighbouring topics Molecular biology stubs, Polymerase chain reaction, Reverse transcriptase inhibitors, so understanding it makes those chapters shorter.
In everyday life: Look for Template-switching polymerase chain reaction outside the textbook — in sport, cooking, traffic, electronics or the sky above you. An example you found yourself is remembered far longer than one you were given.

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How to study Template-switching polymerase chain reaction in 20 minutes

Read the reference excerpt below once, without taking notes.
Close the page and write down what Template-switching polymerase chain reaction means in your own words.
Compare your version with the excerpt and mark what you missed.
Work through the three examples above with pen and paper.
Explain Template-switching polymerase chain reaction out loud to somebody else — or to Teacher Smith in the lgStudy chat.

Frequently asked questions

What is Template-switching polymerase chain reaction in simple terms?

Why does Template-switching polymerase chain reaction matter?

Because it connects several chemistry ideas at once: it gives you a definition you can apply, a quantity you can calculate, and a way to check whether a result is plausible.

How should I study Template-switching polymerase chain reaction?

Read the excerpt, restate it from memory, then work through the examples and applications listed on this page. The five-step study plan above takes about twenty minutes.

What does this page cover?

It gives you a compact reference excerpt plus original lgStudy explanations, examples, applications and study material on Template-switching polymerase chain reaction.