PCR (2 Questions each for 2.5 marks)
Q1. How many primers are needed to amplify copies of a DNA template?
Q2. List the names of the three PCR steps?
Q3. What is the name of the PCR technique where a quantification of DNA product occurs?
Q4. What is the name of the PCR technique where a fluorescent tag is used?
Q5. What is the name of the PCR technique where two sets of primer pairs are used in two successive reactions?
Q3. What is the name of the PCR technique where a number of primer pairs are used to identify various DNA templates in a single reaction?
Chromatography part I
(3 Questions each for 2.5 marks)
- Which phase carries the complex mixture of sample to separate through column chromatography?
- What is the general term used for the phase where the mobile phase flows?
- If we have two components in sample (component A have weaker interactions and component B have strong interactions) which will elute first through column chromatography?
- When we use glass or aluminum sheets as a matrix to hold stationary phase for the separation of components is referred to as which type of chromatography technique?
- Which chromatography technique uses descending technique to hold the stationary phase and matrix?
- If we have two molecules of different sizes (molecule A size is 50 KD and molecule B size is 500 KD) in a sample to be separated through gel filtration chromatography of pore size required to hold 50-100 KD molecules. Which molecule elute early and why?
- If we have mixture of protein macromolecules and we only wants to separate the all negative charge protein then which column chromatography is needful in this separation?
- In water analysis (such as nitrate, phosphate, sulphate, etc.) we commonly use which type of column chromatography?
- Which specific chromatography is helpful when Lectins a class of ligands has been use in the separation and analysis of isoenzymes? Write the name of technique.
Chromatography part II
(3 Questions each for 2.5 marks)
- Write the name of any two chromatography techniques, where gravitational force is required for the mobility of sample?
- What is the main difference between Liquid chromatography (LC) and HPLC?
- Which component is attached in HPLC after the HPLC column?
- What is this image called?
- Which equipment is required to separate and analyze the volatile compounds from serum sample?
- What is called retention time in HPLC?
- What are the ways to identify the concentration of purified compound through HPLC?
- Which type of stationary phase is commonly used in Gas-liquid partition chromatography?
- What is the major difference between column of GC and HPLC?
- What is the role of pneumatic control in GC?
- Why retention of analyte molecules occurs in chromatography?
- What is the typical dimension of capillary column in GC?
- Name any two mobile phase gas uses in GC?
- Which stationary phase is required in GC for Gas-solid chromatography?
- What is the maximum size of most of the analytes in terms of Dalton to be analyzed by GC and why it is prefered?
Mass Spectrometry
(1 Questions each for 2.5 marks)
- Which technique is required to obtain the structural information as well as molecular mass?
- On what basis mass spectrometer analyze the atomic mass of an ion?
ELECTROPHORESIS
(3 Questions each for 2.5 marks)
- Why electrophoresis is different from ion exchange chromatography?
- Why we mostly use agarose gel instead of polyacrylamide gel for DNA analysis through electrophoresis
- Which dye is usually used for visualization of double-stranded DNA on a Gel?
- Why different percentage of gel is prepared to perform electrophoresis?
- Why we use SDS in protein separation through electrophoresis?
- How many amino acids can be combine with one SDS molecule?
- Why Dithiothreitol (DTT) or 2-mercaptoethanol is required in electrophoresis?
- Which disease is defined through this pattern of serum protein electrophoresis?
- Write four major application of SDS-PAGE in clinical diagnosis.