Question

1) You have two human liver cells (A and B) and you hypothesize that the insulin receptor gene in Cell A has a mutation in exon 1 and Cell B contains the wild type sequence.  You extract genomic DNA from each of the cells.  Of the following, what would be the most efficient (quick, precise and relatively cheap) way to test your hypothesis.

a. Isolate protein from both cells, purify the insulin receptor, and determine the amino acid content.

b. Sequence the genomes of both cells using NGS and compare the two sequences.

c. Use RNA-seq to determine the mRNA sequences of both cells

d. Amplify exon 1 of both cells using PCR and determine the sequences using Sanger sequencing

e. Amplify exon 1 of both cells and use qPCR to quantify the amount of PCR product

2) In forensic DNA analysis, multiplex PCR is used to obtain a DNA profile of the CODIS 13 STR loci.  The following is correct

a. The STR loci each contain multiple SNP alleles that are unique to an individual.

b. The CODIS 13 loci have been selected for forensic work because each of the alleles is present at the same frequency in the population.   

c. Multiplex PCR uses 1 pair of PCR primers that are able to amplify all 13 STR loci

d. The PCR products are separated and visualized on an agarose gel

e. Multiplex PCR uses 26 different primers containing fluorescent markers that allow visualization of the size of the PCR products.

3) In order to knock out a sequence in the genome through CRISPR/Cas9 gene editing, all of the following must be present in the cell except:  

a. a plasmid containing the DNA copy (template) of the sequence that you are trying to knock out

b. a source of sgRNA

c. a target sequence within the genome that contains a flanking PAM sequence

d. non homologous end joining (NHEJ) enzymes

e. a source of Cas9 nuclease

Answer 1

1. Answer is option B that is sequencing of both genomes by using NGS that is Next Generation Sequencing and compare that two sequnces, that is efficient and cheap method of determination of hypothesis, rather than other remaining methods.

2. Answer is option e that is multiplex PCR allows 26 different primer containing fluorescent marker that allows visulization of DNA is the correct option.

3. Answer is option d that is non homologus end joining enzymes are not requured in gene editing by CRISPR and Cas9, because it only used when there is breaks in DNA strands.