Genetics Lab Midterm
Terms
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- Law of Segregation
- Homologous chromosome separation during meiosis
- Law of Independent Assortment
- During meiosis the separation of one pair of homologous chromosomes is separate from another pair
- % Accuracy Equation
- (observed/expected) x 100
- Gene
- sequence of DNA that codes for a specific protein
- Allele
- various forms of a gene
- Epistasis
- one gene pair masks the expression of another (CI)
- Parts of the Corn
- Pericarp - outermost layer Aleurone - expressed when ACR pathway is complete Endosperm - double fertilization of 1 sperm and 2 eggs Anthocyanin - pigment that expresses color of Aleurone
- Lab 1 name
- Critical Calculations in biology and practicing with Pipetmen
- Lab 2 name
- Monohybrid and Dihybrid Crosses in Maize
- Lab 3 name
- DNA Isolation
- Lab 4 name
- DNA quantification and PCR
- Lab 5 name
- Gel Electrophoresis and PCR fragment ligation in a plasmid
- Accept or Reject the Null Hypothesis (CHI squared)
- Accept - Xcalc < Xcrit(follows Mendel's Laws) Reject - Xcalc > Xcrit (something other than chance occurred)
- CHI-squared Equation
- (observed - expected)^2 / expected
- If ACR pathway is complete (all three are dominant)
- Anthocyanin Production - Aleurone(Pr) Dominant - purple Recessive - red
- ACR pathway is not complete (at least one is recessive)
- Endosperm color is expressed - Y dominant = yellow y recessive = white
- Sweet corn vs. Feild Corn
- Su dominant = starchy endosperm (smooth) su recessive = high sugar (wrinkled)
- Epistasis in corn
- CI is present - prevents aleurone color (almost like ACR pathway is not complete
- DNAzol purpose
- guanidine detergent lysing solution - breaks lipid membrane, releases DNA
- Absolute ETOH purpose
- causes DNA to condense into a pellet (DNA is hydrophillic - repels ETOH)
- 75% ETOH purpose
- hydrophobic enough to keep DNA in pellet; salts are dissolved
- 8mM NaOH purpose
- dissolves DNA back into solution
- HEPES purpose
- Neutralizes DNA to pH of ~7.5
- Optical Density
- measuring the absorbance of a biomolecule
- Quality Equation
- (260nm reading/280nm reading)
- Optimal DNA/RNA quality
- 1.8/2.0
- Polymerase Chain Reaction purpose
- make a large number of copies of a gene
- PCR Tube
- Add 22.5uL primer mix and (<2.5uL) DNA
- The Cycling Reaction
- Denaturation, Annealing, and Extension
- Denaturation (Cycling Reaction)
- at 94*C - the double stranded DNA forms into single stranded DNA
- Annealing
- at 54*C - sequence is primed so 3'OH connects to 5'phosphate so DNA polymerase can function
- Extension
- at 72*C - bases are coupled to the primer
- Purpose of Gel Electrophoresis
- to separate biomolecules based on size and charge
- Gel Matrix
- Higher % - tighter the matrix
- Ladder
- Estimates the size of the biomolecule 600 - brightest spot 440 - ideal position
- DNA charge
- overall (-) charge; therefore moves towards the (+) end
- TAE Buffer purpose
- contains ions to help conduct a charge
- Ethidium Bromide
- (mutagen) - intercalates between hydrogen bonds on DNA - flourescent under UV light
- Amount of Agarose Needed in Electrophoresis
- .675g of Agarose
- Amount of Buffer needed for Electrophoresis
- 30mL TAE stock and 270mL water
- Restriction Endonuclease
- cut DNA a specific nucleotide sequences leaving unpaired complementary base ends known as sticky ends with can be sealed together with Ligase
- Kaposi's Sarcoma
- associated Herpes virus (KSHV)
- Quality Goal
- to have a higher 260nm than 280nm reading to have a higher overall
- 260nm reading
- detects nucleic acids(DNA)
- 280nm reading
- detects RNA/proteins and nucleic acids - NOT GOOD (Impure)
- Plasmid
- Circular piece of DNA
- Plasmid Cutting
- Ligase anneals cut plasmid to form sticky ends on recombinant DNA
- Reagents for Lab 4
- Primer (mtDNA) - provides 3'OH group dNTP's - provide nucleotides for extension Buffer - enzyme for extension