Title: Exam 3
Course: BTEC-1100-301-302-303-F21 > Section: Module 4 - Applied Molecular Biology

Q: A large number of copies of any DNA segment can be obtained by:
- introducing foreign DNA into a microorganism so that it can be replicated.
- stimulating increased transcription of the appropriate sequence of mRNA.
- stimulating increased translation of the appropriate DNA molecule.
- inducing the production of DNA from proteins.
- stimulating reverse transcription of tRNA.
A: introducing foreign DNA into a microorganism so that it can be replicated.

Q: Splicing together DNA from 2 different organisms is called:
- bioengineering.
- in vitro gene technology.
- biotechnology.
- recombinant DNA technology.
- genetic engineering.
A: recombinant DNA technology.

Q: Transformation is a process whereby:
- bacteria are transferred into plasmid cells.
- viruses are transferred into bacterial cells.
- plasmids are transferred into bacterial cells.
- bacteria are transferred into viral cells.
- plasmids are transferred into viral cells.
A: plasmids are transferred into bacterial cells.

Q: What process or enzyme was used for Step 1 in Figure 14-01?
- PCR
- a bacteriophage
- a reverse transcriptase
- a DNA ligase
- a restriction enzyme
A: a restriction enzyme

Q: In Figure 14-01, the use of antibiotic medium at the end of this process:
- selects against plasmids containing human DNA fragments.
- selects for plasmids containing particular DNA fragments.
- selects for bacteria containing plasmids.
- selects for bacteria lacking plasmids.
- prevents contamination of the medium.
A: selects for bacteria containing plasmids.

Q: ____________ is a technique that can be used to separate DNA molecules on the basis of their size.
- Separation gradient
- Electrophoresis
- Selective sorting
- Cloning
- PCR
A: Electrophoresis

Q: PCR:
- can only be carried out if DNA polymerase is heat-resistant.
- is used to amplify tiny quantities of DNA in vitro.
- only replicates target sequences.
- has applications for archaeology and crime scene analysis.
- All of the above.
A: All of the above.

Q: The chain termination method is used in:
- DNA sequencing.
- gel electrophoresis.
- Southern blotting.
- DNA ligation.
- polymerase chain reaction.
A: DNA sequencing.

Q: The amplification of recombinant plasmids occurs by:
- the growth of the plasmid DNA.
- DNA replication of plasmids outside a host cell.
- the process of growth and division of the host cell.
- transformation of bacterial cells.
- the polymerase chain reaction.
A: the process of growth and division of the host cell.

Q: You need to make stock solutions for the DNA lab class. You need 0.75 molar Tris, 0.25 molar NaCl , and .5 molar DTT. Given the molecular weights as follows;
Tris: 121.1 g/mole
NaCl : 58.0 g/mole
DTT: 154.0 g/mole
How would you prepare 300 milliliters of these stock solutions?

Q: You next want to make 75 milliliters of a buffer containing 1 millimolar DTT, 10 millimolar Tris and 5 millimolar NaCl. From the stock solutions above (i.e.-0.75 M Tris, 0.25M NaCl, 0.5M DTT) how would you prepare this buffer?

Q: Given the following gene sequence in FASTA format, design the forward and reverse primers incorporating the lic site (5'-GGTATTGAGGGTCGC-3') in the forward primer and the lic site (5'-AGAGGAGGTTAGAGCC-3') in the reverse primer. Aim for a 60 degrees C melting temperature.
1500 nucleotides: ANarchY gene
>gi|49175990:400971-402386 Escherichia coli K12, complete genome
ATGAAACAAAGCACTATTGCACTGGCACTCTTACCGTTACTGTTTACCCCTGTGACAAAAGCCCGGACACCAGAAATGCCTGTTCTGGAAAACCGGGCTGCTCAGGGCGATATTACTGCACCCGGCGGTGCTCGCCGTTTAACGGGTGATCAGACTGCCGCTCTGCGTGATTCTCTTAGCGATAAACCTGCAAAAATATTATTTTGCTGATTGGCGATGGGATGGGGGACTCGGAAATTACTGCCGCACGTAATTATGCCGAAGGTGCGGGCGGCTTTTTTAAAGGTATAGATGCCTTACCGCTTACCGGGCAATACACTCACTATGCGCTGAATAAAAAAACCGGCAAACCGGACTACGTCACCGACTCGGCTGCATCAGCAACCGCCTGGTCAACCGGTGTCAAAACCTATAACGGCGCGCTGGGCGTCGATATTCACGAAAAAGATCACCCAACGATTCTGGAAATGGCAAAAGCCGCAGGTCTGGCGACCGGTAACGTTTCTACCGCAGAGTTGCAGGATGCCACGCCCGCTGCGCTGGTGGCACATGTGACCTCGCGCAAATGCTACGGTCCGAGCGCGACCAGTGAAAAATGTCCGGGTAACGCTCTGGAAAAAGGCGGAAAAGGATCGATTACCGAACAGCTGCTTAACGCTCGTGCCGACGTTACGCTTGGCGGCGGCGCAAAAACCTTTGCTGAAACGGCAACCGCTGGTGAATGGCAGGGAAAAACGCTGCGTGAACAGGCACAGGCGCGTGGTTATCAGTTGGTGAGCGATGCTGCCTCACTGAATTCGGTGACGGAAGCGAATCAGCAAAAACCCCTGCTTGGCCTGTTTGCTGACGGCAATATGCCAGTGCGCTGGCTAGGACCGAAAGCAACGTACCATGGCAATATCGATAAGCCCGCAGTCACCTGTACGCCAAATCCGCAACGTAATGACAGTGTACCAACCCTGGCGCAGATGACCGACAAAGCCATTGAATTGTTGAGTAAAAATGAGAAAGGCTTTTTCCTGCA8;X¯ å‚ó‹jAGTTGAAGGTGCGTCAATCGATAAACAGGATCATGCTGCGAATCCTTGTGGGCAAATTGGCGAGACGGTCGATCTCGATGAAGCCGTACAACGGGCGCTGGAATTCGCTAAAAAGGAGGGTAACACGCTGGTCATAGTCACCGCTGATCACGCCCACGCCAGCCAGATTGTTGCGCCGGATACCAAAGCTCCGGGCCTCACCCAGGCGCTAAATACCAAAGATGGCGCAGTGATGGTGATGAGTTACGGGAACTCCGAAGAGGATTCACAAGAACATACCGGCAGTCAGTTGCGTATTGCGGCGTATGGCCCGCATGCCGCCAATGTTGTTGGACTGACCGACCAGACCGATCTCTTCTACACCATGAAAGCCGCTCTGGGGCTGAAATAA

Q: You want to sequence some recombinant DNA. Using two or three short sentences explain how the di-deoxy chemistry and theory works.

Q: Please diagram and explain in several short sentences how the lactose operon works to control gene expression.

Q: Below is the basic scheme for the central dogma. For each step please answer:
Feedback: What the step is called? b) What enzyme is used for each step? c) What important DNA or RNA sequence is necessary?

Q: How would you set up a PCR reaction with the following STOCK solutions;
Forward and reverse primers at 0.8 microgram/microliter
NTP mix with 10 mM dNTPs
5X PCR Buffer
1.5 microgram/microliter Genomic DNA
1.0 unit/ microliter TAQ polymerase
125 mM MgCl2
water

Q: How would you set up an appropriate 50 microliter PCR reaction from these stocks given the following reaction concentrations;
1.5 microgram genomic DNA total
1.5 microgram of each primer total
1X buffer
1.5 mM MgCl2
2 units TAQ polymerase
2.5 mM each NTP

Q: You have your reaction set up. Now show me an appropriate PCR cycling program that you would use for 25 cycles to amplify your gene.