Lab 4 – DNA Purification and RFLP Setup/Simulation

These flashcards cover key concepts related to DNA purification and RFLP analysis from the lab notes.

What is the main objective of Lab 4?

To purify PCR products and cut control regions using restriction endonucleases.

Why is DNA purification necessary in molecular techniques?

To prevent interference from enzymes and buffers which can affect enzymatic reactions.

What are restriction endonucleases commonly used for?

They are used to chop larger DNA molecules into smaller fragments.

How do bacteria utilize restriction enzymes?

Bacteria use them to destroy foreign DNA and protect against viral infections.

What is true about the recognition sites of restriction enzymes?

They are typically 4-6 nucleotides long and palindromic.

What property do palindromic sequences in DNA exhibit?

They have the same sequence on both strands when read 5’→3’.

What is recombinant DNA?

DNA that consists of DNA molecules from different sources combined together.

What is RFLP analysis?

A technique to analyze differences in DNA sequences using restriction enzymes.

How can single nucleotide differences affect RFLP analysis?

They can result in different cutting patterns of DNA, leading to varying fragment sizes on a gel.

What method is commonly used for DNA purification in this lab?

QIAquick PCR Purification Kits.

What are the four major steps involved in the DNA purification method?

Protein denaturation, DNA binding to the column, DNA washing, and DNA elution.

Which restriction enzymes will be used for cutting mtDNA control regions in this lab?

MseI and RsaI.

Why were MseI and RsaI chosen for this experiment?

Because of their variability in recognition sites in human mtDNA.