DNA Polymerase in Lab vs. Cells Flashcards

1
Q

DNA Polymerase in the cell has ________ timed replication of long chromosomes, ______ per cycle, and are done at _______ temperature.

A

precisely, once, same

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2
Q

Because synthesis is 5’ to 3’, replication of complementary DNA strands in the cell is ________ and is ______________.

A

asymmetric, semi-discontinuous

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3
Q

Many ____________ enzymes are required for fast high ________ replication of genomes such as helicase, DNA primase, and DNA ligase.

A

accessory, fidelity

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4
Q

In the lab, _______ _______ reaction amplifies short sequences of interest with steps controlled by ______

A

polymerase chain, temperature

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5
Q

DNA Polymerase active site has _______ , 3’ OH of ______ strand, nucleoside 5’triphosphate, 2Mg+.

A

template, growing

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6
Q

DNA ________ is the stepwise, directional addition of ______ to a growing DNA chain.

A

synthesis, dNTPs

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7
Q

DNA synthesis by polymerases require a _______ strand and a _______.

A

template, primer

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8
Q

DNA strands must be ________ prior to DNA synthesis.

A

separated

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9
Q

DNA polymerization always occurs in the ___ to ___ direction.

A

5’ , 3’

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10
Q

DNA double helix must be denatured for _________. In cells, denaturation is localized and starts at an ______ of replication.

A

replication, origin

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11
Q

A __________ _______ is where DNA replication in cells require many more components than the polymerase.

A

replication fork

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12
Q

The _________ of DNA strands is an ________-_________ process carried out by special enzymes.

A

separation, energy dependent

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13
Q

DNA helicases hydrolyze ____ to induce cyclic conformational changes and mechanical forces which are moving wedges that _____ the helix _____.

A

ATP, pry, apart

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14
Q

_____ _____ ______ proteins bind to the unwound strands of DNA to keep them separate and stop them from forming a helix again.

A

single strand binding

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15
Q

Because the DNA Double Helix is antiparallel, the synthesis of one DNA strand is __________ whereas it’s complementary strand is synthesized in ________.

A

continuous, fragments

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16
Q

DNA polymerases need an ____ primer. DNA primase is an ____ polymerase.

A

RNA, RNA

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17
Q

synthesis of the _______ strand leaves gaps that must be _______ by DNA _______.

A

lagging, sealed, ligase

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18
Q

DNA _______ seals gaps by charging the 5’ phosphate with ____ in the nick, which is subsequently attacked by 3’OH, sealing the DNA with phosphate.

A

ligase, AMP

19
Q

_______ unwind DNA double helix ______ of the replication fork.

A

helicases, infront

20
Q

IN PCR, the template is provided by ______ and can vary widely in ______.

A

you, quality

21
Q

In PCR, DNA primers are provided by _______ and direct where synthesis ________.

A

you, starts

22
Q

In PCR, the DNA strand is separated by ________ but a DNA polymerase that can _________ heat is required.

A

heating, withstand

23
Q

PCR amplifies DNA through successive rounds of _________, _________, and __________ all regulated by temperature.

A

denaturation, annealing, extension

24
Q

PCR results in the ___________ amplification of DNA molecules.

A

exponential

25
primer design requires finding the right _______ temperatures to balance specificity and structure.
melting
26
the melting temp (____) of a primer is the temp at which ___ of the DNA Duplex is denatured into single strand DNA. this reflects the _______ of the DNA sequence.
Tm, 1/2, stability
27
the Tm of primers depend on its ___________. It's best predicted by _____ _________ calculations which account for basepair h bonds and base stacking stabilization.
sequence, nearest neighbor
28
_______ gel electrophoresis separates nucleic acids based on _____.
agarose, size
29
________ transcriptase allows for detection of RNA in addition to DNA
reverse
30
PCR can be used to amplify _________ DNA sequences from complex DNA mixtures.
specific
31
the power of PCR to single out specific DNA molecules can be exploited in the ______
clinic
32
Ex of Clinical use of PCR. 1. take blood sample from infected person 2. remove cells by _________ 3. extract _____ 4. _______ transcription and _____ amplification of particle 5. run sample through _____ _________ with column of uninfected person
centrifugation, RNA, reverse, PCR, gel electrophoresis
33
DNA tells what a cell ____ do, RNA tells what a cell ____ do.
can, does
34
reverse transcriptase is an RNA dependent DNA polymerase; it ______ RNA and ______ DNA
reads, writes
35
reverse transcriptase can be used to make ______ libraries. which are libraries of ________ after post transcriptional processing.
cDNA, mRNA's
36
agarose gel electrophoresis doesnt require ______ as linear DNA molecules naturally have similar _____ and _____ proportional to their size.
SDS, shape, size
37
________ PCR directly detects the exponential growth of PCR products during the PCR in order to _____ specific DNA molecules. an example is the use of fluorophore to know when the strand has ______
quantitative, quantify, binded
38
in quantitative PCR a ________ ______ of known DNA quantities allows you to calculate the original concentration of DNA of known sequence from the ____.
standard curve, Ct
39
amount of DNA is _______ proportional to the cycle number.
inversely
40
________ sequencing is sequencing DNA using dideoxynucleotides (ddNTP) as chain __________. These have a 3' H instead of OH.
sanger, terminators
41
____________ of nucleic acids facilitates easy detection of labeled probe sequences (florescent)
hybridization
42
microarray is an array of synthesized single stranded _____ corresponding to regions of a genome of interest
DNA
43
ChIP + detection - cut DNA into _______ - immunoprecipitate ___ on a DNA binding protein - detect bound ______ -detects where proteins bind
fragments pull DNA