Cell and Molecular Biology - Exam 3 Master Set

Antibodies are ___-shaped, with two ___ domains and one ___ domain. They have 2 ___ chains and two ____ chains

Y, F_Ab, F_C, heavy, light

Antibodies bind to _____ in _____

epitopes, antigens

______ chromatography prepares molecules by specific binding

Affinity

_____ chromatography prepares molecules by their charge

Ion exchange

_____ chromatography prepares molecules by their size

Size exclusion

_____ separates molecules according to their charge and size, using a positive charge that gets applied to a ____

Electrophoresis, gel

______ separates molecules mainly according to their mass by denaturing the protein, using buffers at high pH

SDS-PAGE

The ______ strategy uses a ____ antibody to bind to the protein of interest, and then a _____ antibody to bind to the first one.

Western blots, primary, secondary

Why are secondary antibodies favorable for testing?

easily mass-produced, wastefully coupled to chromogenic enzyme or fluorophore for easy detection, amplification of the signal by multiple binding, saves $ on reagents

______ tests liquid samples directly on a plate where proteins bind to a solid surface. It uses both primary and secondary Ab.

ELISA

In ____ ELISA, the antigen itself is coated straight onto the plate and binds to the primary, then the secondary detects it

standard

____ ELISA is more specific, because it has multiple Abs to “cover all bases'“ of the Ab binding sites on the antigens.

Sandwhich

_____ stains nuclei blue

Hematoxylin

_____ stains basic proteins in the cytoplasm red/pink

Eosin

In Immunofluorescent microscopy, the tissue is preserved with a ______, and then ______ occurs with the addition of a detergent to allow the antibodies to cross into the cell and stain proteins.

PFA fixation, permeabilization

PCRs need oligonucleotide ____ and _____ in order to replicate the DNA.

primers, dNTPs

The key advancement that allowed PCR to be invented was the making of ____ DNApol

thermostable

The process of denaturing and annealing in PCR testing is called _____

thermocycling

In sanger sequencing, the DNA is first denatured so the ____ can bind. DNApol lengthens the strand, and will place a fluorescently labeled ____ randomly, which will terminate the strand. The test continues until all lengths are present.

primer, ddNTP

In NGS, fluorescent ____ are reversible terminators that mark the colors of the nucleotide triplets that make up each gene.

NTPs

Why do we convert RNA into DNA for RNA analysis?

RNA is unstable and rapidly degrades

In order to analyze RNA, we use _____. The template switching is enabled by ______

cDNA, reverse transcriptase

Real Time PCR is based on _____. The fluorophore is only visible when it is _____ from its pair.

FRET, cleaved

RNA-seq is used to quantify the _____ of RNA for every transcript (how much is every gene being expressed)

abundance

In Situ Hybridization fixes the tissue and uses_____ with complementary sequence “barcode” for the gene.

oligonucleotide probe

DNA sequences can also be amplified by growing them in microorganisms. One that we commonly use is _____, because it is a ____

E.coli, plasmid