PCR

Two functions of PCRs?

- specify region of DNA that will be amplified -provide a 3'-OH group which DNA polymerase needs to start synthesizing DNA

necessary "ingredients" for a PCR to proceed.

-Forward Primer - Nucleotides -Reverse Primer - Template DNA -DNA polymerase (or Taq polymerase)

The three main stages of the PCR process are usually repeated around 30 times over several hours. Approximately how many copies of the target region of original DNA molecule are made during that time?

1 billion

DNA polymerase:

an enzyme that can synthesize new DNA in a test tube -Using a heat- stable DNA polymerase allows us to perform repeated rounds of DNA replication without adding any additional DNA polymerase.

The building blocks of DNA are:

nucleotides

PCR stands for

polymerase chain reaction

Short, single stranded pieces of DNA that are designed to base pair (or match up with) a specific segment of DNA you want to copy are called:

primers

PCR Primers:

short pieces of single stranded DNA that are complementary (or matching) to sequences in the DNA extracts.

DNA template:

the DNA that is copied

Deoxyribonucleotides:

the monomers, or building blocks, for DNA. DNA polymerase links deoxyribonucleotides (dNTPs) together into a new DNA strand. All four nucleotides for DNA synthesis are mixed together (indicated by the "N" for any nucleotide in dNTPs).

What is the main purpose of PCR?

to amplify DNA

how big are target regions ?

~ 500-800 bp

If no primers were included in your PCR the reaction would not work because:

Complementary base pairing would not occur in any DNA strands

amplified dna regions are used to create?

DNA barcodes

During this step in PCR, DNA is separated into two single strands.

Denaturation

In sequential order, what are the three steps of PCR?

Denature DNA, Anneal Primers, Extend DNA

Which of the following is NOT a term that can be used for the DNA that you want to make copies of in a PCR?

Expansion DNA (or DNA expansion)

During PCR, DNA polymerase (or Taq polymerase) starts copying at

Primers attached to the end of the desired DNA sequence

PCR CYCLE Steps 1-3

Step 1: Denaturing Step 2: Annealing Step 3: Extension -Cycle repeated ~ 30-40 times, doubling DNA with each cycle

PCR Primers become the starting point for DNA replication (T or F?)

TRUE

If no DNA polymerase (or Taq polymerase) were included in your PCR, the reaction would not work because:

There is no enzyme to make new complementary strands of DNA