Question

If you want to identify genes that change expression levels in a mouse model of liver disease under conditions of stress.

(i) What kind of DNA library will be the best one to use and why?

(ii) Based upon your answer, design an experiment which would allow you to identify genes which change expression in the liver in response to a hepatitis infection. Be sure to include experimental parameters such as sample source, preparation, conditions, methods, controls, analysis, expected results.

Answer 1

Answer(I)c-DNA library will be suitable one to use as it provides the expression of the liver cells, and a large quantity of RNA samples can be used for quantitation.

Answer2) sample source-Male mice and trangenic mouse suffereing from hepatitis, maintained under general condition of light and incubation optimum temperature, RNA extraction and quantitation- the liver cells were isolated and preserved, medium used: The general kits like TRIzol is used, with this ethanol is precipitaed and RNA concentration is maintained and checked for initial purity result 260/280 ratio spectrophotometry, then the samples are quantitated with gene expression analysis-Hepatocytes are maintained in eagles medium and supplemented with glucose,serine,amino acids to provide holistic conditions, further hepatocytes are also treated in presence and absence of staursporine and MTT test and DNA Ladder test is done for evaluation of identification and expression of genes using western Blotting

Expected outcome- With the above mentioned techniques c-DNA microaray,number of genes are expressed under normal (control) and diseases (treated) and helps in analyzing genetic alteration or variations in the sample.