MODERN SYSTEMATIC BACTERIOLOGY

Name the technique used to detect specific proteins from a complex mixture of proteins extracted from cells.

• DNA-DNA hybridization

• Random Amplified Polymorphic DNA (RAPD)

• Southern blot

• Western blot

Select the name of the blotting technique used after separating DNA fragments based on size via electrophoresis.

• Southern blot

• Northern blot

• Western blot

• Eastern blot

Find the best technique to quantify the amplification of targeted DNA molecules.

• Dot-blot hybridization

• Real-time polymerase chain reaction

• Multilocus sequence typing

• Colony hybridization

Choose the technique that exploits variations in homologous DNA sequences by digesting the DNA into fragments using restriction enzymes. These fragments are then separated by gel electrophoresis.

• Amplified Fragment Length Polymorphism (AFLP)

• Multilocus Sequence Typing (MLST)

• Restriction Fragment Length Polymorphism (RFLP)

• Random Amplified Polymorphic DNA (RAPD)

Identify this typing method which differentiates microbes by using primers complementary to the interspersed repetitive consensus sequences and enables the amplification of diverse-sized DNA fragments consisting of sequences between the repetitive elements.

• Restriction Fragment Length Polymorphism (RFLP)

• Repetitive Sequence-based PCR (rep-PCR)

• Random Amplified Polymorphic DNA (RAPD)

• Multilocus Sequence Typing (MLST)

Name the technique that selects bacterial colonies with desired genes.

• Dot-blot hybridization

• Colony hybridization

• Fluorescence in situ hybridization

• Northern blotting

Give the molecular technique used to characterize bacterial isolates by using sequences of internal fragments of seven housekeeping genes.

• Restriction Fragment Length Polymorphism (RFLP)

• Amplified Fragment Length Polymorphism (AFLP)

• Random Amplified Polymorphic DNA (RAPD)

• Multilocus Sequence Typing (MLST)  

This technique requires the extracted DNA to be cut into fragments by using restriction enzyme, and followed by gel electrophoresis in which the electric field periodically changes direction.  Larger pieces of DNA will be slowed down to realign their charge when the field direction is changed, while smaller pieces will migrate quicker. This will separate different fragments of DNA and therefore, different bands will be observed.

• Restriction Fragment Length Polymorphism (RFLP)

• Repetitive Sequence-based PCR (rep-PCR)

• Pulsed-Field Gel Electrophoresis (PFGE)

• Multilocus Sequence Typing (MLST)  

Find the TRUE statement regarding gel electrophoresis.

• DNA will be pulled toward the positively charged end of the gel tank

• The shortest DNA molecules are the nearest to the well where the sample was loaded onto the gel

• Gel electrophoresis separates the mixtures of DNA, RNA or proteins according to molecular size

• The shorter pieces of DNA move through the gel the slowest