0000000000359619
AUTHOR
Maria Cantone
A method for eluting DNA in a wide range of molecular weights from agarose gels
We have developed a simple and rapid method for recovering DNAs of a wide range of molecular weights from agarose gels. A DNA-containing gel slice is placed on a Parafilm sheet in the center of a circular (positive) electrode and covered with a drop of buffer, while a linear (negative) electrode is placed on the top of the gel and driven about 1 mm into the gel itself. When a continuous current is applied, the DNA migrates into the buffer toward the circular electrode. We have obtained almost total recovery of DNAs up to 10 kb in size. Our method may also be used, under appropriate conditions, for higher molecular weight DNAs. The yield and all the biological assays performed on the DNAs ob…
Sequence analysis of the rDNA spacer of Paracentrotus lividus and observations about pre-rRNA processing. NTS sequence of Paracentrotus lividus rDNA.
We have isolated and sequenced one intergenic region and a small part of the flanking regions (18S and 26S rRNA coding regions) of the rRNA-encoding genes (rDNA) from the sea urchin Paracentrotus lividus. This region is about 3.8 Kb long. Northern blot hybridizations and S1 mapping experiments demonstrated the presence of a partially processed 21S rRNA precursor while has the same 5' terminus as the 32S primary precursor, also in developmental stages characterized by a low rate of rRNA synthesis.
Polymorphisms in the intergenic region of the sea urchin Paracentrotus lividus ribosomal DNA
Abstract Blot-hybridizations of the sea urchin Paracentrotus lividus genomic DNA with ribosomal DNA (rDNA) probes revealed individual variations in the length and in the sequence of the non-transcribed spacer (NTS) region. The number of rDNA repeat subclasses distinguishable within any individual sea urchin is usually limited (1 to 3) with respect to the widest polymorphism of the population as a whole. The heterogeneity in sequence is revealed by the presence or the absence of specific restriction sites in the spacer region. The data obtained by the intensity of the polymorphic bands indicate that different mechanisms bring about these two types of polymorphism. Preliminary data also indic…