research techniques

ELISA

a plate based technique used to detect and quantify specific substances such as proteins, antibodies, or hormones

more color = more t______ m_____

target molecules

hybridoma

cell formed by fusing antibody producing B-cell with cancer/myeloma cell so it produces 1 antibody forever

enzyme linked antibody + substrate = _______

creates the color

CRISPR Knockout

a genetic engineering technique that uses CRISPR-Cas9 system to permanently inactivate or silence a specific target gene

What is Cas9?

it is a protein enzyme that cuts DNA at a specific target sequence

What mistake does the repair make after Cas9 cuts the DNA?

non-homologous end joining tries to repair itself but is messy so it makes mistakes called indels which disrupts the gene expression.

flow cytometry

checks individual cells one at a time to see what markers or proteins they have. Usually florescent antibodies bind to the target marker or protein to show if expression positive or negative

forward scatter (FSC)

cell size

Side Scatter (SSC)

cell complexity or gradularity

shRNA Knockdown

cuts mRNA and it is a method to decrease production of a specific protein or decrease the activity of a specific gene

gel electrophoresis

separates DNA, RNA, or proteins by size sign electricity. The smaller the protein/particle, the further it travels. (measures protein size)

western blot

separates proteins, transfers them to a membrane, then uses antibodies to detect one specific protein. (identifies one specific protein)