A cDNA insert [ 9] of human VEGF-C was [P]-dCTP-labeled by the random priming method (Ready-to-go DNA labeling Beads, Amersham Pharmacia Biotech, Piscataway, NJ).
These probes were labeled by the random priming method using the RediPrime II kit (GE Healthcare Uppsala, Sweden) according to the manufacturer's instructions.
The cDNA probes (MUC1, MUC2, MUC5B, MUC5AC, MUC8 and β- actin) were labeled by the random priming technique using DECAprime II kit (Ambion, Austin, TX).
Spelt1, Spelt52, pSc119.2, and pAs1 probes were labeled by the random hexamer method with α-P-dATP (Amersham Pharmacia Biotech, United Kingdom) using Klenow fragment [ 58].
Sample and gender matched reference genomic DNAs (500 ng each) were labeled by the random priming method with fluorescence dyes, Cy 3 and Cy 5, respectively.
A cDNA insert (kindly provided by Professor Kari Alitalo, University of Helsinki, Finland) [ 35] of human VEGF-C was [P]-dCTP-labeled by the random priming method (Ready-to-go DNA labeling Beads, Pharmacia Biotech, Piscataway, NJ).
In the experiment analyzed in this study, as outlined in Fig. 1, each individual CD34+ HSPC is distinctly labeled by the random incorporation of a lentiviral vector in the host genome before transplantation into an animal.
The colonies were then transferred to a Hybond N+ membrane [ 53] and hybridized with the probe labeled by the random hexamer method using α-P-dATP (Amersham Pharmacia Biotech, UK) and a Klenow fragment [ 54].
The probes were labeled by the random-priming method using random primer labelling kit (Genei, Bangalore, India) and northern blotting was carried out.
The immunoprecipitated CpG-methylated DNA (test) and the untreated, sonicated DNA (input control) were labelled by using the random priming with the Nimblegen Dual-Color DNA Labelling kit (Roche-Nimblegen, cat. no. 06370250001) with fluorescent dyes Cy3 and Cy5, respectively.
