Molecular Biology and Genetic Engineering Lecture

These flashcards cover key concepts and details from lectures on molecular biology and genetic engineering, focusing on DNA manipulation techniques, cloning vectors, PCR, and sequencing methods.

What is the benefit of cutting DNA fragments and cloning vectors with the same restriction enzyme?

It creates compatible sticky ends on the two fragments.

What are good features of a cloning vector?

Can replicate in host cell, contains selectable marker or reporter gene, contains sequencing primer sites, contains restriction cut sites.

What is the difference between cloning and transformation?

Cloning means to move a piece of DNA into a vector for amplification, while transformation is inserting foreign DNA into a host cell.

What does the presence of an antibiotic resistance gene in a cloning vector represent?

A selectable marker.

What is the outcome of transforming E. coli with a plasmid containing a kanamycin resistance gene that also has LacZ?

You can observe colonies but can't conclude if they contain the DNA insert without further tests.

When transferring colonies from a kanamycin plate to X-gal, which colonies are chosen?

White colonies, which indicate they have been transformed with the plasmid that contains the DNA insert.

What is a significant advantage of BACs and YACs over plasmids for cloning?

They can hold larger fragments of DNA.

What is a necessary change to the gene of a human enzyme before expressing it in E. coli?

Replace the human promoter with an E. coli promoter and remove the introns.

Which component is NOT part of a standard PCR reaction?

ddNTPs.

Match 95°C in PCR with its corresponding step.

Denaturation - DNA becomes single-stranded.

How does PCR relate to cloning with vectors?

Both generate large copy numbers of a specific segment of DNA.

What process does a Northern blot describe?

Transfer RNA from a gel to a membrane, probe with DNA or RNA molecule.

What is true about next-generation sequencing compared to Sanger sequencing?

NGS utilizes flow cells to bind and analyze multiple DNA molecules at once.

What does transgenic mean in the context of mutation methods?

Gene from one organism is expressed by transforming it into another organism's genome.

What is the role of gRNA in CRISPR-Cas9 gene editing?

It is specific to the gene or DNA region you want to target.