Nano Lab (Lec9 DNA amplication)

Nano Lab (Lec9 DNA amplication) - DNA Amplification...

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Nano Lab MAE C187L/C287L DNA Amplification Techniques Nano Labs, MAE C187L/C287L
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The Central Dogma Replication : The DNA replicates its information in a process that involves many enzymes. Transcription : The DNA codes for the production of messenger RNA (mRNA). Translation : Messenger RNA carries coded information to ribosomes. The ribosomes "read" this information and use it for protein synthesis.
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Nano Lab MAE C187L/C287L What we need is a bait and report When the “red fish” bite
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Nano Lab MAE C187L/C287L Amplification of Target Molecules
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Nano Lab MAE C187L/C287L Polymerase Chain Reaction (PCR) DNA molecules can be mass- produced from incredibly small amounts of material with PCR. Kary Mullis ' discovery allows the chemist to mimic the cell's own natural DNA replication process in a test tube. It has now become much easier to characterise and compare the genetic material from different individuals and organisms.
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Nano Lab MAE C187L/C287L Polymerase Chain Reaction (PCR) PCR is a technique widely used in molecular biology. As PCR progresses, the DNA generated is itself used as template for replication. This sets in motion a chain reaction in which the DNA template is exponentially amplified. With PCR it is possible to amplify a single or few copies of a piece of DNA across several orders of magnitude, generating millions or more copies of the DNA piece.
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Nano Lab MAE C187L/C287L PCR Process 1. DNA to be amplified is put into solution containing short DNA "primers" which can bind to the 3' ends of the DNA, the 4 nucleotide bases: A, C, G, T; and special DNA polymerase (Taq polymerase). 2. DNA is heated to 95 C -> single chains. 3. Solution is then cooled to 50 C; Primers bind to ends of the single strand DNA . 4. The temperature is now raised to 72 C and the DNA polymerase causes the synthesis of new complementary strands to all the single strands This is the end of the first cycle.
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Nano Lab MAE C187L/C287L PCR Process The temperature is cycled from 95 to 55 to 72 over and over. The DNA is doubled at each cycle and at the end of 32 cycles it has been amplified 1 billion times. A cycle can be done in as little as 17 seconds, so it is possible to get a billion- fold amplification in an hour or less including set-up time.
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Rolling circle replication (RCA) describes a process of nucleic acid replication that can rapidly synthesize multiple copies of circular molecules of DNA or RNA, such as plasmids. Some eukaryotic viruses also replicate their DNA via a rolling circle mechanism. Continued DNA synthesis can produce multiple single-stranded linear copies of the original DNA in a continuous head-to-tail series. Rolling Circle Amplification (RCA)
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This note was uploaded on 07/28/2008 for the course MAE 187L taught by Professor Chen during the Spring '08 term at UCLA.

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Nano Lab (Lec9 DNA amplication) - DNA Amplification...

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