PCR and Gel electrophoresis
Polymorphism
Transcription
Translation
Translation
Bacterial
transformation
transformation
Calculations
100
What does PCR stand for?
Polymerase chain reaction
100
(slide) Using PCR & gel electrophoresis, which template and primers could you use to detect an SNP in Exon 1?
Option 1) gDNA, primers L1 and R1
Option 2) cDNA, primers L2 and R2
Neither of these options would work
Both of these options would work
Neither of these options would work
100
What is the name of the step that turns DNA into pre-mRNA and what synthesizes RNA from DNA?
Transcription
RNA polymerase
100
Name the 2 types of promoters and the genes that we used for Experiment 3:
Constitutive: Amp-R and Conditional: GFP
100
How much agarose is required to make a 2% gel with a volume of 300 mL 1X TAE?
6 g
Calculations:
(2g/100) * 300 = 6g
200
What step of PCR involves the separation of DNA strands and what temperature does it require. What is the enzyme involved in PCR?
Denaturation/ 98 C/ DNA polymerase
200
Which SNP mutation changes a codon to a stop codon?
Nonsense
200
This is the term for a three-nucleotide sequence on mRNA that codes for a specific amino acid.
codon
200
What allows the GFP gene to glow?
arabinose
200
If you want to make 5 Liter of 1x TAE from 12.5x TAE buffer, how many mL of TAE are needed?
400 mL
REMEMBER YOUR UNITS
300
What is the name of the DNA Polymerase typically used in PCR and why?
TAQ polymerase because it can survive in high temperatures
300
The mRNA sequence "AUC UAC AGC GAA UAG" codes for the amino acids "I-T-S-G-Stop". The third codon has a SNP mutation that changes the third nucleotide 'C' to an 'A': AGC --> AGA (changing the protein to A) What type of mutation would this be?
Nonsynonymous -SNP
300
What type of RNA serves as a link for growing amino acid chain?
Transfer-RNA
300
How can a bacteria uptake plasmid?
Bacteria is turned chemically competent by weakening their cell walls through chemical washes and heat shock. This allows plasmids to enter the plasmid.
300
If you dilute a solution that has a volume of 300 mL and a concentration of 90 M to make a solution that has a final volume of 500 mL, what will the final concentration be and how much water will you have added?
final concentration = 54M
water volume = 200mL
c1v1 = c2v2
90M * 300mL = c2 * 500mL
final volume =(90*300)/500 = 54M
volume of water = final volume - initial volume
500 - 300 = 200mL
400
(T/F) DNA during gel electrophoresis runs towards the negative charge
False
400
Which of the following polymorphisms would have the most effect: SNP, 1bp insertion, or 3bp deletion?
1bP
Frameshift BAD
400
Where do transcription and translation occur in cell?
Transcription: nucleus
Translation: cytoplasm
400
What is the purpose of the antibiotic resistance gene in plasmids when transforming bacteria?
to select bacteria that have taken up the plasmid
400
Calculate the transformation efficiency: You used 10uL of plasmid at a concentration of 1.5ng/uL to transform into E.coli. You have 500uL of transformed cells, and you plate 200uL onto an LB+Kan plate, which will only grow the transgenic E.coli. You let the plate grow overnight, and observe that 30 colonies grew. What is the transformation efficiency?
5cfu/ng = 5000 cfu/ug
transformation efficiency: (# colonies/ #ng of plasmid) * (#total volume µl/#plated volume)
500
500: Which process is NOT consistent with the central dogma: replication, reverse transcription, or translation?
Reverse transcription
500
In the sequence alignment, where is an Indel present?
(Hint there are 4 answers)
Position 7 (All 4 sequences)
Position 8 (All 4 sequences)
Position 4 (All 4 sequences)
Position 8 (Sequence 2)
Position 6 (All 4 sequences)
Position 5 (All 4 sequences)
Position 3 (All 4 sequences)
Position 7 (All 4 sequences)
Position 8 (All 4 sequences)
Position 4 (All 4 sequences)
Position 8 (Sequence 2)
Position 6 (All 4 sequences)
Position 5 (All 4 sequences)
Position 3 (All 4 sequences)
500
Translate the following mRNA sequence 5’ GAUAAC 3’ in the 5’ to 3’ direction
5’ GUUAUC 3’
500
Transformation efficiency formula?
first calculate mass of plasmid: #ng of plasmid = volume of DNA used * concentration of DNA.
transformation efficiency: (# colonies/ #ng of plasmid) * (#total volume µl/#plated volume)
500
