Quantitative real-time pcr, Biology

1. You are given a pair of primers that has been shown amplify a 125 bp DNA of interest specifically by a end point PCR- as there was only one PCR product with a size of 125 bp detected by agarose gel electrophoresis.  How would you further develop this end-point PCR to be a quantitative real-time PCR?  Please provide a rationale for each step that you choose to further develop the assay and what is the alternative if your first choice is not working.

2.You have been asked to develop a monoclonal antibody against the human His-tagged Casc4 recombinant protein that you purified from the expression construct that you designed earlier in the semester (assume the protein is soluble).

a.) Outline a brief protocol for generating this monoclonal antibody using mice as the host species.

b.) What is a method for purifying this antibody from culture supernatant?

3. Zach wants to analyze the expression of transferrin receptor (a surface transmembrane protein) in an epithelial cell line with and without treatment using a certain drug.
a.) Outline a brief protocol for obtaining these results.
b.) Why did you choose this method?

 

Posted Date: 2/19/2013 8:11:58 AM | Location : United States







Related Discussions:- Quantitative real-time pcr, Assignment Help, Ask Question on Quantitative real-time pcr, Get Answer, Expert's Help, Quantitative real-time pcr Discussions

Write discussion on Quantitative real-time pcr
Your posts are moderated
Related Questions
The acidity and alkalinity (pH) of an environment has a strong influence on the activity and stability of macromolecules such as enzymes. These enzymes play an important role durin

What is the difference between spermatogonium and spermatocyte I? The male germ cells are the spermatogonia (diploid cells, 2n) located in the testicles. They mature and by mea

Explain about the Oncogenic Viruses? Certain viruses that interfere with the functions of the regulatory genes have been identified. These viruses are called oncogenic viruses.

Explain about the Calcium and Hypertension? Chronic inadequate intake of calcium may play some role in etiologies of hypertension. Calcium deficiency has been linked to hyperte

Spermatogenesis - Gametogenesis The procedure of maturation of spermatogonia into sperms starts at puberty (about 14 years) and continues into old age. The spermatogonia that

R i n g w o r m It is also known as dermatophytosis resulting in alopecia, scab formation and ring like lesions in the affected body parts. Eti o logy: Zeop

Q. What are the major cellular features of fungi? There are pluricellular and unicellular fungi. All fungi are heterotrophs and eukaryotes. Fungi have cells with cell wall m

Are fruits always the flesh part of the "fruits"? Is the edible part of the onion a fruit? In some so-called fruits the actual fruit is not the flesh part. For instance, the fl

Inspiration decreases the pressure in the central veins and thus increases the pressure gradient between the peripheral and central veins thus augmenting venous return. Skeletal mu

Air Microbes