Term
| What is the purpose of the Polymerase Chain Reaction? |
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Definition
| To make many copies of a single, specific region from any larger DNA. |
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Term
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Definition
1) Design DNA primers matching the regions you want.
2) When target DNA is single stranded, primers bind, create an accessible 3' end hydroxyl group, allow DNA polymerase to bind and polymerize.
3) At cooler temperatures, the primers anneal to the template DNA by complementary base-pairing.
4) Taq polymerase uses dNTPs to synthesize complementary DNA strand, causing extension from the primer.
5) Repeat cycle of the three steps again, doubling the copies of DNA.
6) Repeat cycle again, up to 20-30 times, to produce millions of copies of template DNA. |
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Term
| What are the elements required for PCR? |
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Definition
| Template DNA, synthesized primers, polymerase, plenty of the four dNTPs. |
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Term
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Definition
- Creating fragments of many lengths.
- Combine: elements of PCR plus ddNTPs.
- Collect DNA strands that are produced. Each strand will end with a ddNTP.
- Use gel electrophoresis to separate the sizes.
- Shorter sequences will travel further in the gel. |
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