Which phosphate in the dNTP should be radioactively labeled?

α-phosphate
For radioactive labeling of DNA, the precursor nucleotide is an [α-32P]dNTP. The phosphate group that is incorporated in the growing nucleic acid chain is the α-phosphate (see Figure 3.7). Figure 3.7. The action of a DNA polymerase showing how a nucleotide triphosphate is added to the primer.

What are α and β and γ?

Alpha denotes the largest particle, and it penetrates the least. Alpha particles carry a positive charge, beta particles carry a negative charge, and gamma rays are neutral. Beta particles are high energy electrons. Gamma rays are waves of electromagnetic energy, or photons.

What is 32P labeling?

32P-postlabeling analysis is an ultrasensitive method for the detection and quantitation of carcinogen-DNA adducts. It has a wide range of applications in human, animal and in vitro studies, and can be used for a wide variety of classes of compound and for the detection of adducts formed by complex mixtures.

Why is it important that one of the dNTPs is radioactively Labelled?

One or more of the dNTPs are radioactively labeled to help visualize the replication products. Finally, each tube gets one of four special nucleotides called dideoxynucleotides (ddNTPs). These nucleotides are recognized and used by DNA polymerase, and can be incorporated at the 3′ end of a growing chain.

What is Alpha 32p?

Guanosine triphosphate, labeled on the alpha phosphate group with 32P, is typically used in reactions involving enzymes that will incorporate the ribonucleoside monophosphate (base, sugar, and alpha-labeled phosphate) into a chain of RNA. Common applications include: RNA labeling. Labeling with: SP6 RNA polymerase.

Why are β β rays are deflected to a greater extent than α α rays?

Beta particles are attracted to the positively charged plate. Beta particles are fast moving electrons with a very low mass and so have a high charge to mass density. They are deflected much more than the heavier alpha particles.

What is Alpha 32P?

What is 32P used for?

Chromic phosphate P 32 is used to treat cancer or related problems. It is put by catheter into the pleura (sac that contains the lungs) or into the peritoneum (sac that contains the liver, stomach, and intestines) to treat the leaking of fluid inside these areas that is caused by cancer.

Why is it important to include a lower concentration of ddNTPs than dNTPs in a sequencing reaction?

Note that the higher the concentration of the ddNTP in the reaction, the shorter the products will be, hence, you will get sequence CLOSER to your primer. With lower concentrations of ddNTP, chain termination will be less likely, and you will get longer products (sequence further AWAY from the primer).

Why is it important to have a radioactively labeled nucleotide in this experiment?

Radiolabeled nucleotides are commonly used for detection of specific nucleic acid sequences. If you are performing an enzymatic assay (phosphorylation with a kinase, end labeling an oligonucleotide with terminal transferase, etc.), it is important to consider how your enzyme is using the nucleotide as a substrate.

What is 32P genetics?

A radioactive isotope of phosphorus, widely used to label nucleic acids; it emits a strong beta particle and has a half-life of 14.3 days. 32. P was used in the famous Hershey-Chase experiment of 1952. See Chronology. From: 32P in A Dictionary of Genetics »