High concentration agarose gel: A new medium for high resolution electrophoresis

Hoch, Hans; Lewallen, Charles G.

doi:10.1016/0003-2697(77)90039-2

Cited by 7 publications

(2 citation statements)

References 3 publications

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“…In a standard experiment, 1-3 fig of RNA dissolved in 8 fiL of water was heated for 2 min at 90 °C and chilled for 2 min on ice. Two microliters of 5 x concentrated buffer L or H was added, and the samples were incubated for 30 min at 0 °C in buffer L or at 37 °C in buffer H. The samples were cooled on ice and loaded on 3% agarose gels (Hoch & Lewallen, 1977) after the addition of 2 fiL of loading buffer containing glycerol. Denatured samples were obtained by incubation at room temperature for 10 min in the presence of 64% formamide and 0.3% formaldehyde.…”

Section: Methodsmentioning

confidence: 99%

A 19-Nucleotide Sequence Upstream of the 5' Major Splice Donor Is Part of the Dimerization Domain of Human Immunodeficiency Virus 1 Genomic RNA

Laughrea¹,

Jetté²

1994

Biochemistry

232

269

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The genome of all retroviruses, including human immunodeficiency virus type 1 (HIV-1), consists of two identical RNAs noncovalently linked near their 5' end. Dimerization of genomic RNA is thought to modulate several steps in the retroviral life cycle, such as recombination, translation, and encapsidation. We report the results of experiments designed to identify the 5' and 3' boundaries of the dimerization domain of the HIV-1 genome: (1) An HIV-1 RNA starting at nucleotide 252 or at other downstream positions (four tested) does not dimerize despite the inclusion of the whole of a previously proposed dimerization domain (nucleotides 295-401); (2) an RNA starting between nucleotides 242 and 249 (five positions tested) dimerizes to a variable extent depending on the starting position; (3) an RNA starting at nucleotide 233 or at other upstream positions (five tested) is fully or > 80% dimeric; (4) an RNA starting at nucleotide 1 but lacking the 233-251 or the 242-251 region is, respectively, fully monomeric or about 50% monomeric; (5) the 343-401 region contains two strings of G's (GGGGG367 and GGG384) that had been postulated to promote genome dimerization through the formation of guanine quartets. We have deleted the 379-401, 358-401, and 343-401 regions from otherwise dimeric RNAs without changing their ability to dimerize. We reach three conclusions: (1) a dimerization signal exists upstream of the major 5' splice donor (nucleotide 290); (2) the previously proposed downstream dimerization domain is insufficient to promote dimerization and has a 3' half that is not necessary to obtain fully dimeric RNAs; (3) the 5' boundary of the HIV-1 dimerization domain is located somewhere between nucleotides 233 and 242, and the 3' boundary is located no farther than at nucleotide 342, making it possible that the 5' and 3' boundaries of the HIV-1 dimerization domain are both located within the leader sequence. We speculate that the 248-270 or 233-285 region forms a hairpin that is the core dimerization domain of HIV-1 RNA.

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Section: Methodsmentioning

confidence: 99%

A 19-Nucleotide Sequence Upstream of the 5' Major Splice Donor Is Part of the Dimerization Domain of Human Immunodeficiency Virus 1 Genomic RNA

Laughrea¹,

Jetté²

1994

Biochemistry

232

269

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“…The gel strength (NIKAN, at 1.5% gel) and the dynamic viscosity were 890 g cm À2 and 22 cps, respectively. Gels of increasing concentrations of agar were prepared by dispersion of appropriate amounts (3,5,9,12,15,20,25 and 30 g) of agar-agar powder in 100 ml deionized water followed by hydrothermal treatment at 145 C. Samples will be referred to as HAn, standing for a Measured by pycnometry (water density ¼ 0.994 g ml À1 , agar-agar (powder) measured density ¼ 1.5 g ml À1 ). b From TGA measurements.…”

Section: Notes and Referencesmentioning

confidence: 99%

High-concentration compact agar gels from hydrothermal synthesis

et al. 2010

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A new111in-bleomycin complex for tumor imaging: Preparation, stability, and distribution in glioma-bearing mice

et al. 1984

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A new 111In-bleomycin complex (111In-BLMC) is here reported. Its radiochemical purity was 99% by thin-layer chromatography (TLC) (Rf 0.65) and in 5% agarose gel electrophoresis in 0.02 M NaHCO3 it migrated toward the anode. Autoradiographs of TLC and gel electrophoresis plates showed no change on storage for 3 weeks. Urine and plasma from untreated or glioma-bearing mice after injection of 111In-BLMC were analyzed by TLC and gel electrophoresis. Results indicated stability in vivo, nonbinding to transferrin, affinity to viable tumor, and excretion faster than 111In-BLM-B2, 111In-BLM, or 57Co-BLM. Tissue distributions 24 hr after injection of radiopharmaceutical showed activity ratios of tumor to blood, muscle, and brain of 13.1, 12.4, and 81.6, respectively, which were significantly higher than those for previously prepared 111In-BLM-B2 or 111In-BLM (except for brain, 0.05 less than P less than 0.1). The new 111In-BLM complex may be useful in clinical imaging and for combining radionuclide radiotherapy and chemotherapy.

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High concentration agarose gel: A new medium for high resolution electrophoresis

Cited by 7 publications

References 3 publications

A 19-Nucleotide Sequence Upstream of the 5' Major Splice Donor Is Part of the Dimerization Domain of Human Immunodeficiency Virus 1 Genomic RNA

A 19-Nucleotide Sequence Upstream of the 5' Major Splice Donor Is Part of the Dimerization Domain of Human Immunodeficiency Virus 1 Genomic RNA

High-concentration compact agar gels from hydrothermal synthesis

A new111in-bleomycin complex for tumor imaging: Preparation, stability, and distribution in glioma-bearing mice

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