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| 1 | |
Match each item in the left column with the most appropriate description in the right column. Each letter in the right column is used once and only once.
| _____ restriction enzymes | A. | Process of creating multiple DNA copies | | _____ methylases | B. | Protect host DNA | | _____ electrophoresis | C. | Molecular scissors | | _____ hybridization | D. | Separates DNA fragments by size | | _____ DNA ligase | E. | Molecular glue | | _____ PCR | F. | Collection of DNA fragments cloned into vectors | | _____ transformation | G. | Induction of bacteria to take up eXogenous DNA | | _____ molecular cloning | H. | Amplification of DNA using DNA polymerase | | _____ library | I. | Base pairing between complementary DNA and/or RNA sequences |
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| | A) | EHAFCBIDG |
| | B) | IGACBDFEH |
| | C) | CBDIEHGAF |
| | D) | GAIFEHCBD |
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| 2 | |
Partial digestion of genomic DNA with a restriction enzyme results in:
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| | A) | fragments with restriction enzyme sites on both ends |
| | B) | fragments with an internal restriction site(s) |
| | C) | cDNA fragments |
| | D) | a & b |
| | E) | all of the above |
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| 3 | |
In reference to the alpha-globin locus or beta-globin locus, the term "locus" means:
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| | A) | a duplicated, ancestral gene. |
| | B) | a chromosome region with a cluster of related globin protein genes. |
| | C) | an alpha-, or a beta-globin protein with a single amino acid change. |
| | D) | the location, in this case red blood cells, where a mutant phenotype is manifested. |
| | E) | a geographical region where a genetic disease like sickle cell anemia is common. |
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| 4 | |
What information is absent from the sequence of a cDNA clone?
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| | A) | intron sequences |
| | B) | promoter sequences |
| | C) | amino acid sequence of the encoded polypeptides |
| | D) | a & b |
| | E) | none of the above |
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| 5 | |
What variable of DNA fragments allows their separation by gel electrophoresis?
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| | A) | charge density |
| | B) | length |
| | C) | base composition |
| | D) | base sequence |
| | E) | introns |
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| 6 | |
From the table below, which combination of enzymes leaves ends that can be ligated together?
| restriction enzyme | recognition site | | 1. BglII | A^GATCT | | 2. EcoRV | GAT^ATC | | 3. Sau3A | ^GATC | | 4. PvuI | CGAT^CG | | 5. BssHII | G^CGCGC |
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| | A) | 2 and 4 |
| | B) | 1 and 3 |
| | C) | 2 and 3 |
| | D) | 3 and 4 |
| | E) | 4 and 5 |
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| 7 | |
A genomic library is made for a haploid organism with a genome of 3 x 109 bp. If you want to clone 5 genome equivalents into a vector library that carries 20 kb per construct, how many individual clones do you need to make?
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| | A) | 3.0 x 104 |
| | B) | 1.2 x 105 |
| | C) | 3.3 x 105 |
| | D) | 7.5 x 105 |
| | E) | 7.5 x 108 |
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| 8 | |
What is the function of the ampr gene in a vector?
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| | A) | To allow resistant transformants to grow in selective medium |
| | B) | To distinguish introns from eXons |
| | C) | To allow viral replication |
| | D) | To screen for vectors with inserts |
| | E) | To allow plasmid replication |
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| 9 | |
The lacZ gene is sometimes included in a cloning vector. What is its purpose?
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| | A) | allow resistant transformants to grow in selective medium |
| | B) | distinguish introns from eXons |
| | C) | allow viral replication |
| | D) | screen for vectors with inserts |
| | E) | allow plasmid replication |
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| 10 | |
A restriction enzyme with a recognition site of CAANNNTTG (where N is any base) cuts an average of every:
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| | A) | 16 bp. |
| | B) | 256 bp. |
| | C) | 400 bp. |
| | D) | 4.1 kb. |
| | E) | 65 kb. |
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| 11 | |
A restriction enzyme with a recognition site of CCPyPyGG (where Py is any pyrimidine) cuts an average of every:
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| | A) | 16 bp. |
| | B) | 256 bp. |
| | C) | 1024 bp. |
| | D) | 4096 bp. |
| | E) | 65 kb. |
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| 12 | |
A circular DNA of 4.7 Mb (Mb=million base pairs) length is cut with a restriction enzyme whose precise recognition sequence is not known. The digest shows ~75 fragments on a pulsed-field gel. What is the most likely conclusion from these data?
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| | A) | The enzyme is a 4-base cutter. |
| | B) | The enzyme is a 6-base cutter. |
| | C) | The enzyme is an 8-base cutter. |
| | D) | The enzyme leaves blunt ends. |
| | E) | This was a partial digest. |
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| 13 | |
Which vector is appropriate for subcloning an insert of 3 kb?
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| | A) | ampr plasmid |
| | B) | YAC |
| | C) | cDNA |
| | D) | cosmid |
| | E) | phage lambda vector |
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| 14 | |
Which of the following statements is false?
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| | A) | If you know the protein sequence, you know the DNA sequence. |
| | B) | If you know the DNA sequence, you can predict a protein sequence. |
| | C) | If you know the recognition sequence, you know the average size of the restriction fragment. |
| | D) | a + c |
| | E) | all are true |
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| 15 | |
You wish to amplify the region in BOLD CAPS below using PCR. Select the correct set of primers. Assume, for the purposes of this question, that an octamer primer is sufficient. (Usually, primers 18-20 nucleotides in length are required).
5'gagatcaggacttaGATTACAGATTACAGATTACAGATTACAggccaagtc3'
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| | A) | 5'AGGACTTA3' and 5'GGCCAAGT3' |
| | B) | 5'TAAGTCCT3' and 5'ACTTGGCC3' |
| | C) | 5'AGGACTTA3' and 5'ACTTGGCC3' |
| | D) | 5'AGGACTTA3' and 5'TGAACCGG3' |
| | E) | none of the above |
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| 16 | |
The diagram below shows the products from the Sanger method of DNA sequencing. What is the sequence of the template strand?
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| | A) | 5'TACAGGT3' |
| | B) | 5'ATGTCCA3' |
| | C) | 5'TGGACAT3' |
| | D) | 5'ACCTGTA3' |
| | E) | 5'TGGACAT3' |
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| 17 | |
The partial amino acid sequence of a protein is: met-gly-trp-cys-ser. A mixture of oligonucleotides containing all possible codons for each amino acid is generated. How many different oligonucleotides are in the mixture?
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| | A) | 12 |
| | B) | 14 |
| | C) | 16 |
| | D) | 32 |
| | E) | 48 |
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| 18 | |
You are subcloning a fragment of genomic DNA into an E. coli plasmid vector. As a first step, you cut out the fragment from an eXisting clone using a restriction enzyme. You then ligate the fragment into a similarly-digested plasmid vector carrying an ampr gene. The site you ligate into is in the middle of the lacZ gene coding for β-galactosidase. After ligation, you transform E. coli with the ligated molecules using a CaCl2 solution or electroporation and plate on X-gal plates with ampicillin. Successful transformation is indicated by ________, while successful insertion of DNA into the vector restriction site is indicated by ________.
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| | A) | ampr colonies; white colonies |
| | B) | white colonies; ampr colonies |
| | C) | ampr colonies; blue colonies |
| | D) | amps colonies, blue colonies |
| | E) | no colonies; white colonies |
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| 19 | |
You want to express human insulin protein in E. coli. Given that you already have a clone containing the pig insulin gene, place the following steps in the proper order:
| a. | probe cDNA library with pig insulin gene clone | | b. | isolate mRNA from human pancreas | | c. | eXpress human insulin in culture | | d. | using reverse transcriptase, make cDNA | | e. | grow up positive clones that hybridize to pig gene | | f. | clone cDNAs into expression vector to make library |
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| | A) | aecbdf |
| | B) | fabdec |
| | C) | abdfec |
| | D) | bdfaec |
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| 20 | |
You are trying to restriction-map a plasmid. An EcoRI digest gives you a 10 kb fragment, an XhoI digest gives you a 2 kb and an 8 kb fragment, and the double digest gives you fragments of 2, 3, and 5 kb. From these results, you deduce that the size of the plasmid is ______ kb, the number of EcoRI sites is ______, the number of XhoI sites is ______, and that there is an EcoRI site within the ______ kb XhoI fragment.
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| | A) | 10; 2; 1; 8 |
| | B) | 10; 1; 2; 8 |
| | C) | 20; 2; 2; 8 |
| | D) | 20; 1; 2; 8 |
| | E) | 10; 1; 2; 2 |
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| 21 | |
You cut a small circular piece of DNA with EcoRI and obtain two fragments of 6 and 4 kb on an agarose gel. You cut the same piece with BamHI and obtain two fragments of 9 and 1 kb. When you perform a double digest, you obtain only 3 fragments (5, 4, and 1 kb). What is the most likely eXplanation?
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| | A) | one of the sites is methylated by the assay conditions |
| | B) | two of the fragments in the double digest are similar in length, thus running as a doublet on the gel |
| | C) | all the BamHI and EcoRI sites overlap |
| | D) | one BamHI and one EcoRI site are very close together |
| | E) | none of the above |
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| 22 | |
You are trying to restriction-map a plasmid and obtain the gel below.
| H | H/B | B | | | | | | | | __ | | | 10 kb | | | | | | | | __ | __ | 6 | | | | | | | | __ | | 4 | | | | | | | | | | | | __ | __ | | 2 |
How many HindIII and BamHI sites are there in this plasmid?
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| | A) | 2 HindIII and 2 BamHI |
| | B) | 2 HindIII and 1 BamHI |
| | C) | 1 HindIII and 2 BamHI |
| | D) | 1 HindIII and 1 BamHI |
| | E) | 2 HindIII and no BamHI |
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| 23 | |
You digest a 20 kb plasmid with EcoRI and obtain fragments of 9 and 11 kb. You digest the same plasmid with HindIII and obtain fragments of 3 and 17 kb. A double digest with EcoRI and HindIII results in fragments of 9, 2, 3, and 6 kb. Which of the following fragments will hybridize to a probe made from the 11 kb EcoRI fragment?
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| | A) | 6 kb EcoRI/HindIII fragment |
| | B) | 3 kb EcoRI/HindIII fragment |
| | C) | 2 kb EcoRI/HindIII fragment. |
| | D) | a & b |
| | E) | all of the above |
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| 24 | |
You digest a 25 kb plasmid with restriction enzymes and obtain the following results:
| enzyme | fragment size (in kb) | | | | | EcoRI | 5, 20 | | BamHI | 12, 13 | | SmaI | 7, 18 | | EcoRI & BamHI | 1, 4, 8, 12 | | BamHI & SmaI | 2, 5, 7, 11 | | EcoRI & SmaI | 3, 5, 7, 10 |
Which fragment will hybridize to the 2 kb BamHI/SmaI fragment?
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| | A) | 7 kb SmaI fragment |
| | B) | 5 kb EcoRI fragment |
| | C) | 12 kb BamHI fragment |
| | D) | 10 kb EcoRI/SmaI fragment |
| | E) | 3 kb EcoRI/SmaI fragment |
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| 25 | |
Which of the following would not have its gene represented in a cDNA library from normal adult red blood cell precursors?
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| | A) | α2 alpha-globin |
| | B) | α1 alpha-globin |
| | C) | ε β-globin |
| | D) | β β-globin |
| | E) | a + c |
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