Match the following terms with their descriptions below. Labeled end fragments a

Question

Match the following terms with their descriptions below. Labeled end fragments are not observed where the protein is bound The binding of a protein to a nucleic acid slows its migration in a gel A poly-dT primer is used to begin synthesis of what will eventually be double strand DNA for cloning Separated on the basis of mass, proteins are probed with specific antibodies Base incorporation results in chain termination A. cDNA library B. Western blotting C. DNase I footprinting D. Gel retardation assay E. Dideoxysequencing

Explanation / Answer

Hi,

1. Dnase1 I foot printing : the DNAse I is an enzyme which binds to the specific regions of the DNA. In order to identify the sequence where it binds foot print assay is carried out. The DNA fragments with and without bound DNAse is analyzed to identify the sequence where it binds.

2. Gel retardation assay : here the protein bound DNA and protein unbound DNA are run on the gel. The protein bound DNA being heavier moves slowly as compared to the need DNA. This is called gel retardation. It helps identify whether the isolated protein has DNA binding domain.

3. cDNA synthesis. cDNA stands for complementary DNA. It is the DNA form of RNA strand. This is created by using random hexadecimal primers which bind to the RNA and convert it to double stranded DNA which is complementary to the original RNA chain.

4. Western blot. This is the technique used to separate proteins based on mass and charge and later identifying them using antibodies or other detection systems.

5. Dideoxysequencing : this is also called as Sanger method of sequencing. Here a Dideoxy, a modified base is added randomly during sequencing. This modified base cannot form bonds with next nucleotide thereby terminating the sequencing. By identifying where there termination has occurred one can know the sequence of the DNA.

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