Introduction to Biotechnology & Recombinant DNA – Key Vocabulary

30 vocabulary flashcards covering fundamental terms and techniques from the lecture on biotechnology and recombinant DNA.

Biotechnology

Any technological application that uses biological systems, living organisms or their derivatives to make or modify products or processes for specific use.

Pharmaceutical Biotechnology

Field that exploits biological systems and modern DNA technologies to manufacture drug substances for human or veterinary medicine.

Recombinant DNA

A DNA molecule formed by joining segments from different sources to create a new genetic combination.

Hybridization (nucleic acids)

Spontaneous annealing of two complementary single-stranded DNA or RNA molecules to form a double-stranded structure.

Complementary Base Pairing

Specific hydrogen-bonding of A with T (or U) and G with C that underlies hybridization and DNA replication.

Probe (molecular biology)

A labeled single-stranded nucleic acid used to detect complementary sequences in samples by hybridization.

Restriction Endonuclease

Bacterial enzyme that recognizes specific short DNA sequences and cleaves the phosphodiester backbone at or near those sites.

Nuclease

Enzyme that cuts phosphodiester bonds within DNA or RNA.

Endonuclease

Nuclease that cleaves internal phosphodiester bonds within a nucleic acid strand.

Exonuclease

Nuclease that removes nucleotides successively from a free end of a nucleic acid chain.

Sticky Ends

Single-stranded 5′ or 3′ overhangs left after restriction enzyme cleavage, facilitating ligation with complementary ends.

Blunt Ends

DNA ends with no overhangs produced when restriction enzymes cut both strands at the same position.

DNA Ligase

Enzyme that joins DNA fragments by forming phosphodiester bonds between adjacent nucleotides.

DNA Polymerase

Template-dependent enzyme that synthesizes DNA strands; requires a primer with a free 3′-OH group.

RNA Polymerase

Enzyme that synthesizes RNA from a DNA template, starting at promoter sequences without a primer.

Promoter

Specific DNA sequence that serves as recognition and initiation site for RNA polymerase during transcription.

Reverse Transcriptase

RNA-dependent DNA polymerase that synthesizes complementary DNA (cDNA) from an RNA template; produced by retroviruses.

cDNA

Complementary DNA synthesized from an mRNA template via reverse transcriptase; lacks introns.

Gel Electrophoresis

Technique that separates charged molecules in an electric field through a gel matrix according to size and shape.

Agarose Gel Electrophoresis

Standard method (0.3–2 % agarose) for separating DNA fragments of ~100–50 000 nucleotides using TBE or TAE buffer.

Polyacrylamide Gel Electrophoresis (PAGE)

High-resolution separation of proteins or small nucleic acids in a polyacrylamide mesh formed by acrylamide and bis-acrylamide.

Bis-acrylamide

Cross-linking agent that links acrylamide chains during polymerization to form polyacrylamide gels.

TEMED

Tetramethylenediamine; catalyst that initiates polyacrylamide gel polymerization.

Urea (in PAGE)

Denaturant added to polyacrylamide gels to keep nucleic acids single-stranded during electrophoresis.

TBE Buffer

Tris-borate-EDTA solution commonly used as running buffer in DNA electrophoresis.

TAE Buffer

Tris-acetate-EDTA solution used as an alternative running buffer for nucleic acid electrophoresis.

Southern Blotting

Transfer of DNA from a gel to a membrane followed by probe hybridization to detect specific sequences.

Northern Blotting

Blotting technique that transfers RNA to a membrane for detection with labeled probes, allowing analysis of transcript levels.

Western Blotting

Membrane-based immunodetection of specific proteins using primary and enzyme-linked secondary antibodies.

Molecular Weight Determination (via gel)

Estimation of nucleic acid or protein size by comparing migration distance in electrophoresis to standards.