Quiz 1 of biotechnology applications

dilution equation

C1V2=C2V2

Weight of solute

%w/v x volume of solution(mL)

Moles of solute

Moles of solute = Desired Molarity(M) x Desired Volume(L)

weight of solute

weight of solute(g) = moles of solute (mol) x Molecular weight (g/mol)

Volume of solute

volume of solute(mL) = weight of solute (g) / Density (g/mL)

covert volume to liter

desired volume / 1000

Molarity (m)

M = moles of solute/volume of solution(L)

Dilution of solutions

m1v1 = m2v2

When Griffith separately injected mice with what

heat-killed bacteria of either strain, the mice lived

what happened when he injected mice with combination

the combination of the heat-killed S-train with live R-strain, mice died

what did Griffith observe when analyzing mice

samples obtained from dead mice injected with the combination, he observed live S-strain bacteria to be present

why is bacteria ideal organisms for transformation

can easily take in exogenous genetic material into their genome and quickly amplify it

bacterial transformation

in natural environment bacteria undergoes bacterial transformation by taking in plasmids from other bacteria ina process call conjugation

Plasmid vectors

Plasmids that are used for experimental purposes to transform bacteria with foreign DNA, laboratory settings scientists artifically create recombinant plasmids

Origin of replication

where replication begins

Antibiotic Resistance Gene

allos the bateria that take in the plasmid to survive on plates in the presence of a certain antibiotic drug, allows competitive growth advantage

multiple cloning site

aids in DNA insertion by containing sites for restriction enzymes to cut the plasmid where the gene of interest can be inserted and ligated

Promoter

drves transcription of the gene of interest

selection marker

usually a fluorescent protein such as a green fluorescent protein (GFP_ or can be an additional antibiotic resistance gene

Gene of interest

Typically codes for the desired protein for expression generally <10kb

Components of Gene Cloning

plasmid, enzymes, selection process

Transformation apporaches

Heat shock, calcium chloride, and electroporation

Necessary

If you take that thing away, thet do tha’t do that thing

sufficient

those sets of things is enough

DNA plasmid

recombinant sequence of GFP is intron, allow protein to be expressed

CRISPR

a set of biological molecules we engineer to damage the molecule where we want them to, most genotoxic is double-stranded, which loses genetic material, we need cas9, repair template, HR macinery

Double-stranded break

could lose genetic material in essential genes, could create harmful mutations, near impossible to repair back to the original

NHEJ

low fidelity, somatic cells

Homolog

break bond of humolog and bind

homologus repair

high fidelity, main form of repair in gametes

Agar

used for preparing solid medium, obtained from seaweeds, has no nutitive value, not affected by the growth of the bacteria, melts at 98C and sets at 42C, 2% agar is employes in solid medium

serial dilution

1.0mL to 9.0 mL = 1:10, 1.mL to 9.0 mL = 1:100

Three major genome editing techniques

fuse DNA-binding “Zinc finger”, TALENs join Foki to the variable DNA-binding domains of bacterial proteins called transcription activator-like effectors, CRISPER-Cas9 which uses components of a prokaryotic immune system

CRISPR origin

first observed in bacteria and later identified in Aacher