triphosphates (dNTPs) are selectively incorporated from a large pool of all four present in the reaction buffer, with the polymerase cleaving off two of each incoming dNTP's phosphate groups as the energy source driving the reaction; each newly incorporated nucleotide in turn presents its 3' hydroxyl for the polymerase to move forward one base and repeat the process on.
Specialists in the US had offered a therapy called deoxynucleoside
But they have been offered the prospect of a new treatment in the US, deoxynucleoside
therapy, which, it is thought, might result in some amelioration of this disease.
5 U/uL, Genscript, Nanjing), 5 uL 10x buffer (Transgene, Beijing), 1 uL 20 mM deoxynucleoside
triphosphate (Transgene), 37 uL of sterile distilled water, 1 uL of each primer (50 uM), and 1 uL of template.
In RT-2 reaction, denatured dsRNA was then added to the reaction mixture consisting of 10 l of the 5X reaction buffer, 1 l of 10 mM deoxynucleoside
triphosphate (10mM dNTP) mixture, 2 l of 25 mM MgSO4, 1 l of MuLV Reverse Transcriptase enzyme (5U/ul), 1 l of Taq Polymerase (5U/ul) and 3 l of dimethyl sulfoxide in a final volume of 50 l and then preincubated at 480C for 45 min followed by 35 cycles of 940C for 1 min.
01% gelatin), and 200 mmol/L each deoxynucleoside
triphosphate (dNTP) and 1.
2 mM of each deoxynucleoside
triphosphate (Bioline, UK).
Amplifications were performed in 25ul reaction mixtures consisting of the following: 100 pmol of each primer, 200 uM each deoxynucleoside
triphosphate (dNTP), 10 mM Tris-HCl (pH 8.
2 pM (each) deoxynucleoside
triphosphate (dATP, dGTP, dTTP and dCTP) from CinnaGen-Iran, and 1X PCR reaction buffer 1.
7 Ci/mmol) was obtained from PerkinElmer[R], deoxynucleoside
triphosphates obtained from Promega, Taq polymerase and Moloney leukemia virus reverse transcriptase were purchased from Invitrogen and RNeasy minikit was purchased from Qiagen.
5mM each of deoxynucleoside
triphosphate and 1 U [micro][L.
For each PCR reaction a 25L reaction mixture was prepared which included 20 pmol of each primer 100M deoxynucleoside
triphosphate 10X reaction buffer (100 mM Tris-HCl 15 mM MgCl2 50 mM KCl 0.