Hybrid isolation by recovery of RNA-DNA hybrids from agar using S1 nuclease.
نویسندگان
چکیده
A method for recovering RNA-DNA hybrids from agar employing a single strand specific nuclease is described. The procedure is suitable for large scale isolations, and immobilization of the DNA in agar prior to hybridization allows a high yield of hybrid without interference by DNA reannealing.
منابع مشابه
Assay of DNA-RNA hybrids by S1 nuclease digestion and adsorption to DEAE-cellulose filters.
A fast and accurate assay procedure for DNA-RNA hybrids is described in which exhaustive digestion of unhybridized DNA with S1 nuclease is followed by binding of hybrids to filter discs of DEAE-cellulose. The digested DNA can be efficiently washed from the filters so that background levels of 0.1-0.2% of input tracer DNA can be achieved, in contrast to the much higher (approximately 1-5%) backg...
متن کاملA putative precursor for the small ribosomal RNA from mitochondria of Saccharomyces cerevisiae.
We have characterized a putative precursor RNA (15.5S) for the 15S ribosomal RNA in mitochondria of Saccharomyces cerevisiae. Hybrids were formed with mitochondrial RNA and mtDNA fragments terminally labelled at restriction sites located within the gene coding for 15S ribosomal RNA and treated with S1 nuclease (Berk, A.J. and Sharp, J.A. (1977) 12, 721-732). Sites of resistant hybrids were meas...
متن کاملAlkaline agarose gel electrophoresis.
INTRODUCTION Alkaline agarose gels are run at a pH that is sufficiently high to denature double-stranded DNA. The denatured DNA is maintained in a single-stranded state and migrates through the alkaline gel as a function of its size. Alkaline agarose gels are used chiefly to measure the size of first and second strands of cDNA (Construction of cDNA Libraries Stage 1: Synthesis of First-strand c...
متن کاملSearch for nucleic acid sequences complementary to a murine oncornaviral genome in poly(A)-rich RNA of human leukemic cells.
The presence of viral-like sequences in the RNA of various types of leukemic cells was investigated by hybridizing cellular poly(A)-containing RNA with cDNA synthesized in an endogenous system of purified Moloney murine sarcoma virus [M-MSV-(MLV)]. Poly(A)-RNA-cDNA hybrids were detected by assaying their resistance to S1 nuclease. Hybrids were found in 22 out of the 46 leukemias that were teste...
متن کاملDetecting differential expression of parental or progenitor alleles in genetic hybrids and allopolyploids.
Three assays useful for detecting specific RNA transcripts are primer extension, S1 nuclease protection, and reverse-transcription-cleaved amplified polymorphic sequence (RT-CAPS) analysis. All three of these techniques are used routinely for gene expression analyses and allow insights not possible by RNA blot (northern blot) hybridization. In this chapter, we describe how the primer extension,...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Nucleic acids research
دوره 1 6 شماره
صفحات -
تاریخ انتشار 1974