L reaction volume with 1 [micro]L of deoxynucleoside
triphosphate mix (12.
The reaction was performed by PTC-200 programmable thermocycler (MJ research, USA) in a 20 [micro]l mixture consisting of 15 U avian myeloblastosis virus (AMV) reverse transcriptase, 20 U of RNasin ribonuclease inhibitor, 25 ng random primers, 5 mM MgCl2, 1 mM deoxynucleoside
triphosphate (dNTP) mix and 1X reverse transcription buffer (10 mM Tris-HCl at pH 8.
25U Taq DNA polymerase (Professional Biotech, New Delhi), 200 [micro]M each of the four deoxynucleoside
triphosphates, (1 [micro]l) 100 pmol of each oligonucleotide primer of bla SHV, and (1 [micro]l) 30 pmol of each oligonucleotide primer of [bla.
5mM magnesium chloride, 200 [micro]M each of deoxynucleoside
triphosphates (deoxyadenosine triphosphate, deoxycytidine triphosphate, deoxyguanosine triphosphate, and deoxythymidine triphosphate), 0.
Malaysia), 200 [micro]mol/L deoxynucleoside
triphosates, and 1.
Reverse transcription of total RNA (1 [micro]g) was performed by using hexamer primers, deoxynucleoside
triphosphates (dNTPs), and Maloney murine leukemia virus (MMLV) reverse transcriptase.
Samples were added to a PCR mixture containing a final concentration of 200 mmol/L (each) deoxynucleoside
triphosphate mixture, 10X Hot Start PCR Buffer (Fermentas/Thermo Scientific, Vilnius, Lithuania), 0.
2+], and biotinylated deoxynucleoside
triphosphates (dNTPs) in TdT buffer were added to cover the sections and incubated in a humid atmosphere at 37[degrees]C for 60 minutes.
where dNMP is deoxynucleoside
monophosphate, and [P.
4 U KlenTaq1[TM] (Ab Peptides) with 64 ng anti-Taq antibody (eEnzyme), and 200 [micro]mol/L of each deoxynucleoside
2], 250 [micro]M of each deoxynucleoside
triphosphate (dNTPs) and 2.
5 [micro]g) from Zi geese, 500 ng/[micro]l of random hexamers primer (Promega, USA), 10 mM deoxynucleoside
triphosphate (dNTP) Mix (Takara) and sterile MilliQ water (to a total volume of 12 [micro]l) were heated to 65[degrees]C for 5 min in order to disrupt possible secondary structures and then quickly chilled on ice.