DNA Manipulation

Enzymes, PCR, and Gel Electrophoresis for Biotechnology

Probes

________ are single- stranded DNA or RNA molecules that are labeled with a radioactive or fluorescent marker.

Southern blotting

________ is a technique used to detect specific DNA sequences in a complex mixture of DNA fragments.

Short tandem repeats

________ (STRs) are repetitive DNA sequences that are found in specific locations in the genome.

Methylene blue

________ is a dye that binds to DNA, making the fragments more visible.

buffer

The running ________ provides the ions needed to carry the electrical current during electrophoresis, as well as maintaining the pH and preventing the gel from overheating.

Polymerase

________ is an enzyme that synthesizes new DNA strands by adding nucleotides to the 3 'end of a primer.

forensic analysis

They are used in ________ to identify individuals based on their unique STR profiles.

gel electrophoresis

The fragments are separated by ________ and transferred to a membrane.

Primers

________ are short DNA sequences that bind to specific regions of the template DNA during PCR.

equipment

The ________ used to perform PCR typically includes a thermal cycler, pipettes, and PCR tubes or plates.

Ligase

________ is like glue that seals the gaps that polymerase makes by making new bonds.

Prokaryotic DNA

________ is circular therefore one cut produces one fragment.

plasmid map

A(n) ________ is a diagram of the plasmid showing the location of different DNA sequences and the location of restriction sites as well as base pair sizes.

semilog graph

A(n) ________ is used to estimate the size of DNA fragments based on their migration distance in a gel.

dye

Loading ________ is used to help visualize the samples during electrophoresis and to ensure that they sink into the wells.

molecular biology

They are used in ________ to cut DNA into fragments that can be analyzed, manipulated, or inserted into other DNA molecules.

Electrophoresis safety

________ includes wearing gloves, avoiding contact with the buffer solution, and turning off the power supply before opening the electrophoresis chamber.

PCR

________ (polymerase chain reaction) is a technique used to amplify a specific segment of DNA, making millions of copies in a relatively short amount of time.

extension stage

In the ________, a DNA polymerase adds nucleotides to the 3 'end of the primers, extending the new strands.

Sticky cuts

________ leave overhangs that are complementary to another strand, while blunt cuts produce fragments with a straight end.

DNA ladder

The ________ is a mixture of DNA fragments of known sizes that is used as a standard for determining the size of unknown.

Restriction enzymes

________ are enzymes that cut DNA at specific sequences, called restriction sites.

DNA

________ has a negative charge due to its phosphate groups, and it migrates towards the positive electrode during electrophoresis.

Gel electrophoresis

________ is a technique used to separate and analyze DNA fragments based on their size and charge.

DNA ladder

To read a gel, you can determine the number and size of DNA fragments based on their position in the gel relative to the ________.

The stages of PCR are

denaturation (at 94-98°C), annealing (at 50-65°C), and extension (at 72°C)

Gel Electrophoresis