| |
FIG. 5. Blot hybridization (Northern analysis) of electrophoretically
separated RNA from Xenopus tissues. Poly(A)'
RNA (5 pg) from the indicated tissue was subjected to electrophoresis
on a 1.5% agarose gel as described under "Experimental Procedures"
and blotted onto a nylon membrane. Hybridization was carried out
at 37 "C for 16 h with a mixture of three single-stranded, uniformly
'"P-labeled Xenopus LDL receptor 1 cDNA probes of 150-200 nucleotides
in length (5 X lo6 cpm/ml). The primers for the three :'2P
probes corresponded to nucleotides 482-502, 1318-1338, and 2180-
2197 in pXLDLR-1. The primers were extended on an M13 template
in the presence of [a-'*P]dCTP, the products were subjected to
electrophoresis, and the bands corresponding to the longest products
were eluted from the gels and mixed together. After blotting, the
RNA-containing filter was exposed to Kodak XAR-5 film with an
intensifying screen for 5 h at -70 "C. The positions of RNA standards
run in an adjacent lane are indicated. |
