Methods for northern blot
have been described previously (23).
Southern blot and Northern blot
hybridizations were performed as described previously by Lee et al.
Image bands from Western blot, Northern blot
, and PCR/qPCRs were quantified using Image J software (NIH website).
In order to assess cell binding and internalization of the selected aptamers without the need for labeling, we performed Northern blot
One empty vector control and five Ubi-hpCP:Hc-Pro primary transgenic lines (T0) were analyzed for transgene copy number by Southern blot hybridization and for the accumulation of siRNAs by Northern blot
Sensitive and specific detection of microRNAs by northern blots
analysis using LNA- modified oligonucleotide probes.
For Northern blot
analysis, total RNA was extracted from leaves using Trizol reagent (Invitrogen), and hybridizations were carried out with [[[alpha].sup.32] P] dCTP-labeled PHYB probe.
Nitrocellulose membranes for Western blot and nylon membranes for Northern blot
were purchased from Invitrogen (Carlsbad, CA) and NEN Life Science Products (Boston, MA), respectively.
Transgene expression was first analysed in back skin, brain, and liver of all transgenic lines by northern blot
using a [sup.32]P-labelled oligonucleotide A probe and a U6 RNA (a ubiquitously expressed component of the spliceosome) probe for loading normalization.
Small RNAs were isolated using the mirVana miRNA isolation kit (Ambion Austin, USA) and analyzed by Northern blot
hybridization according SequaGel Sequencing system Kit (EC-833) (National Diagnostics Atlanta, USA) according to manufacturer's instructions.
Expression patterns of cPdcl2 during testicular and ovarian development in chickens were examined by quantitative real-time PCR (qRT-PCR), Northern blot
hybridization, and in situ hybridization.