A virtual pyrogram for hypothetical DNA sequence ACGT, where the polymerase chain reaction product bottom-strand with annealed top-strand sequencing primer is shown on the left, shaded box, and the extended nascent strand after addition of deoxyadenosine
triphosphate (dATP) and deoxycytidine triphosphate are shown in the right, shaded box.
deoxynucleotide triphosphates (dNTPs), which are the nucleotides deoxyadenosine
triphosphate (dATP), deoxyguanosine triphosphate (dGTP), deoxycytidine triphosphate (dCTP), and deoxythymidine triphosphate (dTTP) that compose DNA.(2)
In the disease, the lack of ADA enzyme allows the buildup of 2' deoxyadenosine
triphosphate, a chemical that is particularly detrimental to immune system cells, especially T cells.
DCK is an enzyme that phosphorylates purines (deoxyadenosine
and deoxygaunosine) and a pyrimidine (deoxycytidine) as well as their analogues used as chemotherapeutic agents.
Chinese hamster ovary (CHO) cells were cultured in [alpha]-MEM supplemented with adenosine, deoxyadenosine
, thymidine (0.2 mg/mL each), and 10% FBS.
TA cloning is achieved by Taq DNA polymerase which has non-template-dependent terminal transferase activity which helps adding of a single deoxyadenosine
(A) to the 3' ends of PCR products.
The reaction master mixes for both primer sets had the following similarities: Each was adjusted to a final volume of 95 [micro]L using high-pressure liquid chromatography-grade water and contained 2.5 IU of AmpliTaq Gold DNA polymerase (Applied Biosystems, Streetsville, Ontario, Canada), 1X PCR buffer II (Applied Biosystems), 1.5 mM magnesium chloride, 200 [micro] Meachof deoxyadenosine
triphosphate, deoxycytidine triphosphate, deoxyguanosine triphosphate, and thymidine monophosphate.
This process undergoes deoxyadenosine
methylase (Dam) dependent methylation of GATC sequence in E.
In turn, Cytochrome-c engages apoptotic protease activating factor 1 (APAF 1) and forms the apoptosome in the presence of deoxyadenosine
Because of the substitution of the hydrogen atom with a chlorine atom in position 2 in the deoxyadenosine
ring, cladribine is resistant to ADA action.
Ions were monitored in selected ion-monitoring mode at m/z 256 and 258 for adenosine and 198 and 200 for deoxyadenosine
; these represent the [M.sub.+0], [M.sub.+2] mass isotopomers of the dominant ion after ionization.
The first PCR was performed in 50 [micro]L reaction volume containing 20mM Tris (pH 8.3), 50mM potassium chloride, 1.5mM magnesium chloride, 200 [micro]M each of deoxynucleoside triphosphates (deoxyadenosine
triphosphate, deoxycytidine triphosphate, deoxyguanosine triphosphate, and deoxythymidine triphosphate), 0.5[micro]M each of primers, 0.25 units of AmpliTaq polymerase (GIBCO-BRL, Life Technologies, Gaithersburg, Md), and 5 [micro]L of the digested sample, which contained extracted DNA from at least 50 cells.