Molecular Biology Chapter 23

Front 1

polymerase chain reaction (PCR)

Back 1

Amplification of a DNA sequence by repeated cycles of strand separation and replication.

Front 2

PCR primers

Back 2

Short pieces of single-stranded DNA that match the sequences at either end of the target DNA segment and which are needed to initiate DNA synthesis in PCR

Front 3

target sequence

Back 3

Sequence within the original DNA template that is amplified in a PCR reaction.

Front 4

PCR machine or thermocycler

Back 4

Machine used to rapidly shift samples between several temperatures in a pre-set order (for PCR)

Front 5

Taq polymerase

Back 5

Heat resistant DNA polymerase from Thermus aquaticus that is used for PCR.

Front 6

Thermus aquaticus

Back 6

Thermophilic bacterium found in hot springs and used as a source of thermostable DNA polymerase

Front 7

degenerate primer

Back 7

Primer with several alternative bases at certain positions

Front 8

inverse PCR

Back 8

Method for using PCR to amplify unknown sequences by circularizing the template molecule

Front 9

randomly amplified polymorphic DNA (RAPD)

Back 9

Method for testing genetic relatedness using PCR to amplify arbitrarily chosen sequences

Front 10

TA cloning

Back 10

Procedure that uses Taq polymerase to generate single 3'-A overhangs on the ends of DNA segments that are used to clone DNA into a vector with matching 3'-T overhangs

Front 11

TA cloning vector

Back 11

Vector with single 3'-T overhangs (in its linearized form) that is used to clone DNA segments with a single 3'A overhangs generated by Taq polymerase

Front 12

complementary DNA (cDNA)

Back 12

Version of a gene that lacks the introns and is made from the corresponding mRNA by using reverse transcriptase

Front 13

reverse transcriptase

Back 13

Enzyme that starts with RNA and makes a DNA copy of the genetic information

Front 14

reverse transcriptase PCR (RT-PCR)

Back 14

Variant of PCR that allows genes to be amplified and cloned as intron-free DNA copies by starting with mRNA and using reverse transcriptase

Front 15

differential display PCR

Back 15

Variant of RT-PCR that specifically amplifies messenger RNA from eukaryotic cells using oligo (dT) primers

Front 16

anchor sequence

Back 16

Sequence added to primers or probes that may be used for binding to a support or may incorporate convenient restriction sites, primer binding sites for future manipulations, or primer binding sites for subsequent PCR reactions

Front 17

molecular sewing

Back 17

Creation of a hybrid gene by joining segments from multiple sources using PCR.

Front 18

overlap primer

Back 18

PCR primer that matches small regions of two different gene segments and is used in joining segments of DNA from different sources

Front 19

rapid amplification of cDNA ends (RACE)

Back 19

RT-PCR-based technique that generates the complete 5' or 3' ends of cDNA sequence starting from a partial sequence

Front 20

directed mutagenesis

Back 20

Deliberate alteration of the DNA sequence of a gene by any of a variety of artificial techniques

Front 21

fluorescence resonance energy transfer (FRET)

Back 21

Transfer of energy from short-wavelength fluorophore so quenching the short wave emission

Front 22

SYBER Green I

Back 22

A DNA-binding fluorescent dye that binds only to double-stranded DNA and becomes fluorescent only when bound

Front 23

TaqMan probe

Back 23

Fluorescent probe consisting of two fluorophores linked by a DNA probe sequence. Fluorescence increases only after the fluorophores are separated by degradation of the linking DNA.

Front 24

molecular beacon

Back 24

A fluorescent probe molecule that contains both a fluorophore and a quenching group and the fluoresces only when it binds to a specific DNA target sequence

Front 25

Scorpion primer

Back 25

DNA primer joined to a molecular beacon by an inert linker. When the probe sequence bind target DNA, the quencher and fluorophore are seperated allowing fluorescence

Front 26

quencher

Back 26

Molecule that prevents fluorescence by bind to the fluorophore and absorbing its activation energy

Front 27

rolling circle amplification technology (RCAT)

Back 27

Method based on rolling circle replication that uses DNA polymerase to amplify target DNA at normal temperatures