1 Flashcards

(33 cards)

1
Q

Transfer to a solid support

A

Element two of WB

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2
Q

USed in research to separate and identify proteins

A

Western blot

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3
Q

Separation by size

A

Element one of WB

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4
Q

Marking target protein with antibodies

A

Element three of WB

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5
Q

Prevents antibodies from binding to membrane nonspecifically

A

Blocking

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6
Q

Confirm the identity of the protein and activity of the antibody

A

positive control

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7
Q

Null cell line used to confirm that the staining is not nonspecific

A

negative control

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8
Q

Slightly acidic with lower acrylamide concentration to make a porous gel to separate proteins poorly but allows them to form thin, sharply defined bands

A

Stacking gel

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9
Q

Basic with high polyacrylamide making the gels pores narrower

A

Separating gel

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10
Q

USes electric field oriented perpendicular to the surface of the gel causing proteins to move out of the gel and onto the membrane

A

Transfer

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11
Q

High affinity for protein and its retention abilities. Brittle and doesn’t allow the membrane to be used for reprobing

A

Nitrocellulose membrane

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12
Q

Important for minimizing background and removing unbound antibodies

A

Washing

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13
Q

Detects the protein of interest, Monoclonal or polyclonal

A

Primary antibody

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14
Q

Binds to conserved regions on the primary antibody and acts to amplify the signal, increasing detection sensitivity. Labeled with either an enzyme for colorimetric or chemiluminescent detection

A

Secondary antibody

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15
Q

A chemical reaction in which an enzyme catalyzes the oxidation of a chemical substrate, and the reaction produces light as a byproduct.

A

Chemiluminescent

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16
Q

Primary or secondary antibody labeled with fluorescent molecule excitable by light

A

Fluorescence detection

17
Q

RNA is transcribed into complementary DNA by reverse transcriptase from total or messenger RNA. cDNA is used as a template for qPCR reaction

18
Q

RT and PCR are performed separately with optimized buffers, reaction conditions, and priming strategies

A

Two step RT-qPCR

19
Q

OFten used due to disadvantages for template of RT such as fewer purification steps to ensure more quantitative recovery, better normalization, and it avoids enrichment steps

20
Q

Standardized nuclei extraction by sonication, followed by intranuclear enzymatic chromatin cutting and barcode ligation

21
Q

Assay for transposable accessible chromatin with high-throughput sequencing, a method for mapping chromatin accessibility genome wide

22
Q

Quantify DNA concentrations from multiple samples by analyzing fluorescent signal intensities that are proportional to the amount of amplicon after completing chip assay and sample purification

23
Q

Starts with crosslinking of DNA protein complexes, then samples are fragmented and treated with exonuclease to trim unbound oligonucleotides. Protein-specific antibodies are used to immunoprecipitate the DNA-protein complex. DNA is extracted and sequenced

A

ChIP-seq basis

24
Q

Better mechanical support and allows the blot to be reprobed and stored. Background is higher and requires careful washing

A

PVDF membrane

25
Enzyme used to amplify the signal in photometric assays through catalyzing the conversion of chromogenic or chemiluminescent substrates for the detection of targets
Harseradish peroxidase
26
Combines RT and PCR in a single tube and buffer using RT along with DNA polymerase. Utilizes specific primers
One step Rt-qPCR
27
More sensitivity for template of reverse transcriptase
mRNA
28
Stretch of thymine residues that anneal to the poly A tails of mRNA
OligodTs
29
Enzyme that makes DNA from RNA. Want a high thermal stability
Reverse transcriptase
30
1. Predenaturation if RNA template has a high degree of secondary structure 2. Primer extension 3. cDNA synthesis makes complement DNA strand to the mRNA template 4. Reaction termination prevents qPCR inhibition by active reverse transcriptase
cDNA synthesis
31
Barcoding method to enable high throughput ChIP_seq using common molecular biology techniques. Restriction enzyme-based labeling of chromatin in situ
RELACS
32
Identifying genome wide DNA binding sites for transcription factors and other proteins
ChIP-seq
33
DNA binding dye that drastically increases fluorescence when bound to double-stranded DNA
SYBR green