Techniques in Molecular Biology

Extract plant DNA using the CTAB method by grinding leaf tissue in liquid nitrogen, releasing DNA, then precipitating it and removing proteins, lipids, and phenols.

Explore how electrophoresis moves charged particles in a gel under an electric field. Compare gel electrophoresis with polyacrylamide gel electrophoresis and learn its DNA, RNA, and protein analysis applications.

Quantify DNA using a spectrophotometer, assess concentration at 260 and 280 nm, and purity with an A260/A280 near 1.8 for dsDNA; discuss forensic, paternity, and medical applications.

Learn how agarose gel electrophoresis separates DNA and RNA by size and charge, using pore size control, visualization with ethidium bromide, and DNA ladder-based size estimation.

Learn how SDS-PAGE uses sodium dodecyl sulfate to denature proteins and give a uniform negative charge for size-based separation in separating and stacking polyacrylamide gels.

Explore nucleic acid hybridization in this module, covering primers, probes, blotting methods, and detection in DNA, RNA, and proteins, including PCR microarrays with radiolabeled, enzyme-labeled, or fluorescent labels.

Learn the southern blotting workflow to detect specific DNA sequences within a smear, using restriction digestion, gel electrophoresis, membrane transfer, probe hybridization, and autoradiography.

Learn northern blotting to detect RNA and measure gene expression per the central dogma. Transfer RNA from an agarose gel to a nylon membrane and probe with labeled oligonucleotides.

Discover western blotting to detect a protein of interest from a mixture, using SDS-PAGE, electrophoretic transfer, blocking, antibody probing, and chemiluminescent detection for disease diagnosis and proteome study.

Learn how PCR amplifies DNA with a thermal cycler, DNA template, forward and reverse primers, Taq polymerase, dNTPs, and buffers through denaturation, annealing, extension cycles, and gel electrophoresis.

Design primers 18–30 nucleotides long with 50–60% GC content and unique sequences to ensure specific amplification; verify PCR products by electrophoresis and note applications like genetic fingerprinting and prenatal testing.

DNA sequencing determines the sequence of nucleotide bases in DNA using a sequencing primer, terminator bases, fluorescently labeled bases detected by a laser, and DNA polymerase to extend the strand.

Learn how capillary electrophoresis reads terminator-labeled bases to convert color sequences into DNA sequences, and explore sequencing applications from genomes and disease genes to paternity, agriculture, evolutionary biology, and metagenomics.

Learn key molecular biology components and roles—buffer, liquid nitrogen, gels and stains, DNA ladder, blotting types (southern, northern, western), DNA, RNA and protein detection methods.

Learn the PCR flowchart and core steps: denaturation, annealing, extension, then compare PCR with RT-PCR, including starting material, reverse transcriptase, cDNA, and applications like gene expression studies and COVID-19 detection.

Compare agarose gel and PAGE in biomolecule separation: agarose suits DNA and large fragments with lower resolution, horizontal runs; PAGE gives higher resolution for DNA and proteins with vertical runs.

Explore blotting techniques in molecular biology, including Southern, Northern, and Western blotting for DNA, RNA, and proteins, with HIV detection and genetic disorder analysis.