Polymerase cycling assembly

Polymerase chain reaction

Restriction endonuclease digestion

Gibson assembly

DNA ligation

The start codon should be 5' to the biochemical tag and the stop codon should be 3' to the protein of interest.

The start codons should be 5' to the biochemical tag and between the tag and the protein of interest. The stop codon should be 3' to the protein of interest.

The start codon should be between the biochemical tag and the protein of interest. The stop codon should be 3' to the protein of interest.

The start codon should be 5' of the biochemical tag and the stop codon should be between the tag and the protein of interest.

The start codon should be 5' to the biochemical tag and the stop codons should be between the tag and the protein of interest as well as 3' to the protein of interest.

Two restriction enzymes are used to cut both the plasmid and the subject DNA to be incorporated

Use of alkaline phosphatase to create nicks in the ends of a cut plasmid, preventing ligation by ligase

A cloning reaction that uses a marker system like the lacZ marker to identify recombinant plasmids

All of these

None of these

The ampicillin in the nutrient agar plates expired

The plasmid does not confer ampicillin resistance to the bacteria

The heat shock transformation did not successfully introduce plasmid DNA into the bacteria

The plasmid mutated and now expresses eGFP constitutively

There is not enough information given to interpret the results

Introduce pores in the membrane

Denature the plasmid DNA

Stimulate Escherichia coli proliferation

Mask the negative charge of phosphate groups on DNA

Linearize the plasmid

All of these steps are necessary

None of these steps are necessary

Subject DNA is annealed to sticky ends in the phage

A plate of bacteria is infected with the phage

The phage is specially engineered to insert DNA when it is a recombinant phage

The optimal temperature at which primers bind specifically to their complementary sequences

The optimal temperature at which DNA polymerase is active

The temperature at which the template DNA denatures

Annealing temperatures do not apply to polymerase chain reactions

The temperature at which nucleotides are incorporated into the newly synthesizing DNA strand

Calcium chloride surrounds the bacterial membrane and attracts negatively charged DNA

Calcium chloride mechanically permeates the plasma membrane to allow in genetic material

None of the other answers

Calcium chloride facilitates plasma membrane restructuring following passage of genetic material into the cell

Calcium chloride promotes bacterial colony growth on agar plates following transformation

None of these

Restriction digest the bacterial gene from genomic DNA, PCR amplify the vector backbone from genomic DNA, ligate the gene and vector backbone

Ligate the bacterial gene and vector backbone, PCR amplify the bacterial gene from cDNA, restriction digest the vector backbone

Restriction digest the bacterial gene from cDNA, restriction digest the vector backbone, PCR amplify the bacterial gene and vector backbone together

Restriction digest the vector backbone from genomic DNA, restriction digest the bacterial gene, ligate the bacterial gene and vector backbone

All of these

None of these

Capillary chromatography

Four different fluorescent DNA tags

Thermostable polymerases