Final Flashcards
0.2 µl
P-2
250µl
P-1000
2.5
P-20
25µl
P-200
What is incorrect about gel electrophoresis?
- Gel electrophoresis alone can be used to detect SNPs
- DNA fragments that are very small appear at the top of the gel
What volumes of stock and water are needed to make 500mL of a 50mM solution starting with 200mM stock?
0.125L of stock, 0.375L water
These bind to the template strand of DNA during the anneal step of PCR
Primers
This synthesizes RNA from DNA
RNA Polymearse
Located in the cytoplasm, these use rRNA to conduct translation
Ribosomes
This removes introns
Spliceosome
This gets sent to the splicesome for splicing
pre- mRNA
Included in the MyTaq, these provide the code for DNA replication
dNTPs (Deoxynucleotide triphospahates
Located in the cytoplasm, this contains the DNA code (exons) that will be used to make proteins
mRNA
This makes up the cDNA, containing the information needed for translation
Exons
When pre-mRNA is processes to mRNA, these are removed
Introns
What volume of stock and water is required to make a 250mL of a 15% solution starting with 30% stock
125mL of stock, 125mL of water
What is the (processes, products, organelles) of genetic information transfer with the correct location it occurs in the cell (for eukaryotes)
Nucleus
1.
2.
3.
4.
5.
Cytoplasm
1.
2.
3.
4.
5.
Nucleus
1. Splicing
2. pre-mRNA
3. DNA
4. Transcription
5. Splicesome
Cytoplasm
1. Translation
2. Nucleus
3. tRNA
4. Ribosome
5. Processed polypeptide (protein)
Practice PCR table
Place the orders of magnitude in largest to smallest
Highest
1.
2.
3.
4.
5.
6.
Lowest
Highest
1. Kilo
2. 1
3. milli
4. micro
5. nano
6. pico
Lowest
Dr. Burnette has Glofish in his office that express genes that make them fluorescent (glow certain colors). The red Glofish expresses the Red Fluorescence Protein (RFP). Bacteria were transformed to contain a plasmid with RFP and subsequently plated and grown on three types of media and the phenotype was observed after a period of incubation.
Plate UV light results
LB + Tet + Galactose No fluorescence
LB + Tet + Lactose Red fluorescence
LB + Tet + Sucrose No fluorescence
What is the Activator/Inducer that controls the promoter
Lactose
What type of promoter controls the RFP?
Conditional
Why is water added to the Master Mix?
To dilute the concentration of the working solution
Inherited DNA polymorphisms exists because:
Errors in DNA replication occur naturally during meiosis
Why are a specific type of cell used in Bacterial Transformation (promoter activation experiment)?
They are more capable of taking up plasmids
What further processing needs to occur to pre-mRNA before translation as mRNA?
Removal of introns via splicing
If you dilute a solution that has a volume of 210 mL and a concentration of 75 M to make a solution that has a final volume of 525 mL, what will the final concentration be and how much water will you have added?
30M, and 315L of water
What is incorrect about transcription?
DNA polymerase transcribes gDNA into pre-mRNA
Which of the following statments about setting up a PCR reaction is incorrect?
- Template for all samples should be added to the master mix before aliquoting into strip tubes
- After all components are added to create the mastermix it should not be mixed
- The C1V1=C2V2 equation should be used to determine what volume of MyTaq is needed for the reaction.
- The number of reactions of master mix prepared should equal the number of samples
- Template for all samples should be added to the master mix before aliquoting into strip tubes
- After all components are added to create the mastermix it should not be mixed
- The number of reactions of master mix prepared should equal the number of samples
What is the name of the type of cells that were used during the promoter activation experiment that allowed them to take up the plasmid DNA easier?
Competent
What is false about translation?
- Ribosomes and tRNAs are key components of translation.
- The end product of translation is a chain of nucleotides.
- Translation is one of the steps required for gene expression as protein.
- mRNA is the template for translation.
- The end product of translation is a chain of nucleotides.
What step of the PCR reaction involves primers binding to the DNA template strands?
Annealing
Explain steps of PCR
- Denaturing 98 degrees (breaks double helix into one single strand)
- Annealing 50-70 degrees (primers bind to complementary nucleotides)
- Extension 72 degrees (extending primers from 5’ to 3’ direction)
How much agarose is required to make a 3% gel with a volume of 300 mL 1X TAE? [Hint: 1% gel = 0.5g in 50 mL TAE]
9g
Nymphia needs to make 50 mL of diluted TAE solution for
her experiment. The prep room has 100 mL of 1X (M) TAE
stock solution. What concentration of TAE will Nymphia
need to prepare?
2X
We want to make a 2.4% gel
2.4% represents 2.4g per 100mL of TAE
If we only want 50mL of our gel mixture, then to solve for the amount of Agarose we need
1.2
