5 [micro]mol/L of each deoxynucleotide
triphosphate (50% Cy5-labeled dCTP and dTTP; Amersham Biosciences), 2x extension buffer, 0.
D, A complementary primer (5"-AAGGGT-3') has been annealed, and if the complementary deoxynucleotide
triphosphate (dNTP) is added (dCTP), the DNA polymerase (gray oval) will incorporate it into the nascent elongating strand at the position of the open box.
8 [micro]M each) in a 50-[micro]L mix containing deoxynucleotide
A DNA template is mixed with a thermostable DNA polymerase such as Mg, free deoxynucleotide
triphosphate molecules (dNTPs), and short oligonucleotides called primers, which are complementary for the beginning and end of the sequence to be amplified.
That molecular material is in the form of deoxynucleotide
triphosphates (dNTPs), which are the building blocks for DNA.
triphosphates (dNTPs, the building blocks of nucleic acids: adenine, guanine, cytosine, and thymine), short oligonucleotide primers (needed to initiate DNA synthesis), a thermostable DNA polymerase (that can withstand temperatures in excess of 94[degrees]C experienced during the PCR reaction), and some template DNA have been added to the tubes, DNA replication can be carried out through the use of a thermocycler (see Figure 1A).
Fluorescent terminal deoxynucleotide
transferasemediated dUTP nick-end labelling (TUNEL) was performed on 5 Am-thick transverse sections of sample A.
Genomic DNA (~ 50 ng) was incubated in a total reaction volume of 50[micro]l containing equal concentration of the forward primer 5' TCC CTG AGG GCA TGA GGC T-3' and reverse primers--5' TGA GGG TCA CAC AGG TTC CT-3' (-70 picomoles) (GenScript Corp, USA), 200 [micro]M deoxynucleotide
triphosphate, 10X PCR buffer pH-8.
For the detection of apoptosis the terminal deoxynucleotide
transferase end labelling (TUNEL) method was used according to the instructions given by the supplier.
Research in our laboratory has demonstrated enhanced cellular invasion in MDA-231 breast cancer cells by ODN-362, a 25-base (CpG) deoxynucleotide
6 mM MgCl2, 640 deoxynucleotide
triphosphate, buffer 1x, 0.
In addition, terminal deoxynucleotide
transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL)-positive cells are colocalized with FasL in MS lesions.
At 24 hr after UVR, DNA photoproducts [cyclobutane pyrimidine dimers (CPDs) and 6-4 photoproducts (6-4PPs)] and apoptosis were measured using the enzyme-linked immunosorbent assay and the two-color TUNEL (terminal deoxynucleotide
transferase dUTP nick end labeling) assay, respectively.
5 [micro]l), and four deoxynucleotide
triphosphates (200 mM each), primers (20 pM each), and 0.