Lab Skills
I am have learn new lab techniques and skills during my time at WSSU. In my Molecular Biology and Biotechnology classes I master new lab techniques such as replica printing, DNA and RNA extraction, Western and Southern Bolts, PCR and TLC.
Replica Printing
Replica plating is a microbiological technique in which one or more secondary petri plates containing different solid (agar-based) selective growth media are inoculated with the same colonies of microorganisms from a primary plate (or master dish). The technique involves pressing a velveteen-covered disk, and then imprinting secondary plates with cells in colonies removed from the original plate by the material. Generally, large numbers of colonies (roughly 30-300) are replica plated due to the difficulty in streaking each out individually onto a separate plate.
DNA Extraction and Southern Blot
DNA isolation is essential for genotypic analysis.
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the first step is to lyse the cells to extract to the DNA.
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different solution were used to isolate DNA according to the protocol used.
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2X ethanol is used to precipitate the DNA.
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DNA is stored in the TE buffer
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DNA can be quantified using the Nano-drop or electrophoresis.
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Next, the DNA in the electrophoresis gel was transfer to a membrane through Southern Blot.
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Upright capillary transfer method is used to transfer the DNA to the membrane
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The Southern Hybridization is used to study how genes are organized within genomes by mapping restriction sides in and around segments of DNA
My Channel
Polymerization: Nylon 6.6
Nylon is a linear poly-amide, nylon, is prepared by the step-growth condensation polymerization of adipoyl chloride with 1,6-hexanediamine.
An interfacial polymerization is used to prepare nylon as fibers.
PCR (Polymerase Chain Reaction)
PCT is a revolutionary method developed by Kary Mullis in the 1980s. PCR is based on using the ability of DNA polymerase to synthesize new strand of DNA complementary to the offered template strand. Because DNA polymerase can add a nucleotide only onto a preexisting 3'-OH group, it needs a primer to which it can add the first nucleotide. This requirement makes it possible to delineate a specific region of template sequence that the researcher wants to amplify. At the end of the PCR reaction, the specific sequence will be accumulated in billions of copies
Thin Layer Chromatography
Thin-layer chromatography (TLC) is a chromatography technique used to separate non-volatile mixtures Thin-layer chromatography is performed on a sheet of glass, plastic, or aluminium foil, which is coated with a thin layer of adsorbent material, usually silica gel, aluminium oxide (alumina), or cellulose. This layer of adsorbent is known as the stationary phase.
After the sample has been applied on the plate, a solvent or solvent mixture (known as the mobile phase) is drawn up the plate via capillary action. Because different analytes ascend the TLC plate at different rates, separation is achieved. The mobile phase has different properties from the stationary phase. the TLC is used to detect the completion of the reaction. It is also used to check the purity of the compound.
Above pictures of TLC plate were taken at 5, 30 and 45 mins under the UV light. SM stands for starting material and Rx is the reaction product.