BIO203 Midterm Current Papers Fall 2022

Mcqs: · (Mostly from Files) · Short Questions: (Most Ans are Taken from PPTs) · (Repeated) · 1. dNTPs

Deoxynucleoside triphosphates (dNTPs) are the building blocks from which the DNA polymerase synthesizes a new DNA strand during successive cycles of PCR

dNTPs consist of four basic nucleotides - dATP, dCTP, dGTP, and dTTP.The concentration of each dNTP (dATP, dCTP, dGTP, dTTP) in the PCR is usually 200 μM.

2. Nested and Multiplex PCR

3. In situ PCR

 In Situ PCR (ISH) is a polymerase chain reaction that actually takes place inside the cell on a slide.  In situ PCR amplification can be performed on fixed tissue or cells.

 Applies the methodology of hybridization of the nucleic acids.

 Allows identification of cellular markers  Limited to detection of non-genomic material such as RNA.

4. Type 1 of Restriction and Modification (R-M)

Type I

•  Type I systems were the first to be characterized from E. coli K 12
•  The active enzyme consists of two restriction subunit, two modification subunit and one recognition subunit
•  Type I systems are of little value for gene manipulation

5. Negative control

 Controls for specificity of the amplification reaction contains all reagents and a DNA template lacking the target sequence.  The PCR product should be of the expected size.

6. Nomenclature of The Smith and Nathan's (1973)

•  A suitable system was proposed by Smith and Nathans (1973)
•  The species name of host organisms is identified by the first letter of genus and first two letters of specific epithet
•  E. coli = Eco
•  H. influenzae = Hin
•  Strain identification is written as Eco K
•  In case, host strain has several restriction and modification systems, these are identified by roman numerals, for example, in case of H. influenzae
•  HindI , HindII

7. dNTPs or Assembly PCR: · Deoxynucleoside triphosphates · (dNTPs) · ASSEMBLY PCR

Deoxynucleoside triphosphates (dNTPs) are the building blocks from which the DNA polymerase synthesizes a new DNA strand during successive cycles of PCR

dNTPs consist of four basic nucleotides - dATP, dCTP, dGTP, and dTTP.The concentration of each dNTP (dATP, dCTP, dGTP, dTTP) in the PCR is usually 200 μM.  Assembly PCR used to assemble two or more pieces of DNA into one piece. Assembly PCR PRINCIPLE

 Assembly PCR is the synthesis of long DNA structures by performing PCR on a pool of long oligonucleotides with short overlapping segments, to assemble two or more pieces of DNA into one piece.

8. Application of PCR in microbiology.

 FUNGUS

9. Steps of PCR

THREE STEPS  Denaturation

 Annealing  Extension S