Noise-free chemiluminescent detection of human T cell receptor and interleukin-2 receptor genes after optimization of digoxigenin labeled probe concentration.
Reduction in the background problems to improve the efficiency of nonradioactive labeling and detection procedure has been the focus of attention in the recent past for wider acceptance of the technique in nucleic acid research. We have achieved success in obtaining a relatively background free detection of single copy genes such as, T-cell receptor-delta (TCR-delta) and interleukin-2 receptor (L-2r) in human genome by using optimized concentrations of the digoxigenin labeled probes. Inclusion of such an optimization step for each probe before carrying out the actual hybridization experiment did not require any further modification in hybridization conditions and detection protocols as suggested earlier [Anal Biochem, 210 (1993) 235; Colloq Boehringer Mannheim, 2 (1991) 4.]
Sachdeva
G
G
School of Life Sciences, Jawaharlal Nehru University, New Delhi, India.
Kaur
G
G
Bamezai
R
R
D015139:Blotting, Southern; D004076:Digoxigenin / Q000737:chemistry*; D006801:Humans; D008163:Luminescent Measurements; D015335:Molecular Probes; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000235:genetics*; D015375:Receptors, Interleukin-2 / Q000235:genetics*
D015335:Molecular Probes; D016692:Receptors, Antigen, T-Cell, gamma-delta; D015375:Receptors, Interleukin-2; D004076:Digoxigenin