Separation of complementry strands of plasmid DNA using the biotin-avidin system and its application to heteroduplex formation and RNA/DNA hybridizations in electron microscopy

Delius, Hajo; Heerikhuizen, Harm van; Clarke, Jill; Koller, Barbara

doi:10.1093/nar/13.15.5457

Cited by 26 publications

(3 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Separated single strands of the H cDNA plasmids were prepared by incorporation of biotinylated dUTP into the termini of the EcoRI-linearized plasmid DNA, followed by a second cut by SalI restriction enzyme, and the fractionation of the complementary strands of the two fragments on an avidin agarose column (26).…”

Section: Electron Microscopy Analysismentioning

confidence: 99%

Structure of gene and pseudogenes of human apoferritin H

Costanzo¹,

Colombo²,

Staempfli³

et al. 1986

Nucleic Acids Research

126

View full text Add to dashboard Cite

Ferritin is composed of two subunits, H and L. cDNA's coding for these proteins from human liver (1,2,3), lymphocytes (4) and from the monocyte-like cell line U937 (5) have been cloned and sequenced. Southern blot analysis on total human DNA reveals that there are many DNA segments hybridizing to the apoferritin H and L cDNA probes (1,2,4,6). In view of the tissue heterogeneity of ferritin molecules (7,8), it appeared possible that apoferritin molecules could be coded by a family of genes differentially expressed in various tissues (1,2). In this paper we describe the cloning and sequencing of the gene coding for human apoferritin H. This gene has three introns; the exon sequence is identical to that of cDNA's isolated from human liver, lymphocytes, HeLa cells and endothelial cells. In addition we show that at least 15 intronless pseudogenes exist, with features suggesting that they were originated by reverse transcription and insertion. On the basis of these results we conclude that only one gene is responsible for the synthesis of the majority of apoferritin H mRNA in various tissues examined, and that probably all the other DNA segments hybridizing with apoferritin cDNA are pseudogenes.

show abstract

Section: Electron Microscopy Analysismentioning

confidence: 99%

Structure of gene and pseudogenes of human apoferritin H

Costanzo¹,

Colombo²,

Staempfli³

et al. 1986

Nucleic Acids Research

126

View full text Add to dashboard Cite

show abstract

“…However, if secondary structures are present in the oligonucleotide, high-quality purification is difficult. Numerous techniques already exist for DNA amplification of oligonucleotides through enzymatic synthesis, such as cloning [ 11 ], PCR [ 12 ] and rolling circle DNA synthesis. The latter method, the so-called circle-to-circle amplification (C2CA) [ 13 ], is a method for the amplification of a single stranded DNA sequence, based on successive rounds of ligation, rolling circle DNA synthesis and restriction digestion.…”

Section: Introductionmentioning

confidence: 99%

A new enzymatic route for production of long 5'-phosphorylated oligonucleotides using suicide cassettes and rolling circle DNA synthesis

2007

View full text Add to dashboard Cite

Background: The quality of chemically synthesized oligonucleotides falls with the length of the oligonucleotide, not least due to depurinations and premature termination during production. This limits the use of long oligonucleotides in assays where long high-quality oligonucleotides are needed (e.g. padlock probes). Another problem with chemically synthesized oligonucleotides is that secondary structures contained within an oligonucleotide reduce the efficiency of HPLC and/or PAGE purification. Additionally, ligation of chemically synthesized oligonucleotides is less efficient than the ligation of enzymatically produced DNA molecules.

show abstract

“…Chemical groups introduced into nucleic acid probes have also been used as affinity labels to isolate sequences of interest by hybridization followed by affinity chromatography on solid matrices. Biotinylated DNA in conjunction with avidin matrices (12,20,21), mercurated DNA with thiol matrices (22,23) and poly(A)-tailed RNA with an oligo(dT) matrice (24) are examples of affinity pairs used for isolation of specific DNA fragments.…”

Section: Introductionmentioning

confidence: 99%

Fast quantification of nucleic acid hybrids by affinity-based hybrid collection

1986

View full text Add to dashboard Cite

A hybridization technique for the quantification of nucleic acids is described. In the method a probe pair is allowed to form hybrids with the target nucleic acid in solution. One of the probes has been modified with an affinity label, by which the formed hybrids can be isolated after the reaction. Streptavidin-agarose was used to capture hybrids containing biotinylated DNA. The hybrids were measured using radioiodine as label on the second probe. The rate of the hybridization reaction in solution is fast, allowing the whole procedure to be carried out in 3 h. The method is quantitative with a detection limit of 4 X 10(5) molecules (0.67 attomoles) target DNA. The test is insensitive to impurities in biological samples, which are analyzed without purification of the target DNA. Non-isotopic measurement of the hybrids can also be applied. In this case the hybrids are bound to microtitration wells and detected spectrophotometrically by peroxidase-catalyzed colour development.

show abstract

Separation of complementry strands of plasmid DNA using the biotin-avidin system and its application to heteroduplex formation and RNA/DNA hybridizations in electron microscopy

Cited by 26 publications

References 12 publications

Structure of gene and pseudogenes of human apoferritin H

Structure of gene and pseudogenes of human apoferritin H

A new enzymatic route for production of long 5'-phosphorylated oligonucleotides using suicide cassettes and rolling circle DNA synthesis

Fast quantification of nucleic acid hybrids by affinity-based hybrid collection

Contact Info

Product

Resources

About