Question 1

When cloning a gene into a plasmid vector, two common issues are (i) inability to control the gene's orientation and (ii) vector self-ligation. Which simple solution addresses both problems?

Accepted Answer

Use two different restriction enzymes to generate non-compatible sticky ends.

Question 2

Which plasmid feature determines the number of plasmid copies maintained in a bacterial cell?

Accepted Answer

Origin of replication

Question 3

In CRISPR-Cas genome editing, the guide RNA (gRNA) is engineered by combining two natural RNAs. During editing, which two molecules must the gRNA bind to?

Accepted Answer

Cas9 protein and target DNA

Question 4

A researcher wants to determine which bacterial species are present in an environmental water sample. Which technique is most appropriate?

Accepted Answer

Metagenome construction

Question 5

Which research technique would you use to find well-studied protein motifs encoded by a new gene of interest?

Accepted Answer

BLAST search

Question 6

To identify non-coding regulatory sequences shared by several related bird species, which technique should be used?

Accepted Answer

Phylogenetic shadowing

Question 7

Which technique is best for determining which proteins bind to a protein of interest?

Accepted Answer

Yeast two-hybrid

Question 8

A scientist wants to determine which gene promoters are bound by a putative transcription factor. Which technique should they use?

Accepted Answer

Chromatin immunoprecipitation sequencing (ChIP-seq)

Question 9

Which technique would best identify genes critical for yeast growth in a specific condition?

Accepted Answer

Barcoded knockout library

Question 10

To analyze gene expression in a specific cell type of a multicellular organism, which technique is most suitable?

Accepted Answer

RNA sequencing (RNA-seq)

Question 11

A researcher wants to annotate protein-encoding genes in the genome of a newly discovered insect species. Which technique should they use?

Accepted Answer

Pangenome construction

Question 12

Which technique is best for identifying genetic determinants indicative of a rare genetic disorder using thousands of sequenced genomes?

Accepted Answer

Genome-wide association study (GWAS)

Question 13

What unique information can a translational reporter provide about a gene that a transcriptional reporter cannot?

Accepted Answer

Intracellular localization of the gene product

Question 14

You have chemically mutagenized yeast and found an interesting phenotype. What is the best approach to identify the gene mutation responsible?

Accepted Answer

Sequence the genome

Question 15

Which statement is accurate regarding insertional mutagenesis compared to chemical mutagenesis?

Accepted Answer

You can more easily identify where mutations are in the genome than with chemical mutagenesis.

Question 16

A yeast mutant grows poorly on galactose. After further mutagenesis, you find a second mutation in a separate gene that restores normal growth. What type of genetic screen have you performed?

Accepted Answer

Suppressor screen

Question 17

You want to see how gene expression changes when an organism is subjected to a specific growth condition. Which research technique is best?

Accepted Answer

RNA sequencing (RNA-seq)

Question 18

To determine where histones with a specific H3K9Ac modification bind in the genome, which technique should you use?

Accepted Answer

Chromatin immunoprecipitation sequencing (ChIP-seq)

Question 19

A researcher is using CRISPR-Cas9 to edit a gene in human cells. If the target DNA sequence is $$5'	ext{-}GGGCGTACCTGAGTCCGAA-3'$$, and the PAM sequence is $$5'	ext{-}NGG-3'$$, which of the following is a valid PAM for Cas9 binding?

Accepted Answer

$$5'	ext{-}CGG-3'$$

Question 20

Suppose you want to knock out a gene in yeast and confirm the knockout by PCR. Which combination of primers would best distinguish between wild-type and knockout alleles?

Accepted Answer

Primers flanking the insertion site, so that the PCR product size differs between wild-type and knockout.

Genetics Exam Study Guide: Cloning, CRISPR, Genomics, and Functional Genetics

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