UV Spectrophotometric Analysis of Ribonucleic Acids

Rapley, Ralph; Heptinstall, John

doi:10.1385/0-89603-494-1:65

Cited by 6 publications

(3 citation statements)

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“…Integrity of the isolated RNA was verified by the consistent presence of two sharp bands (corresponding to 28S and 18S rRNA) in agarose/formaldehyde gels electrophoregrams stained with SYBR Gold (Molecular Probes, Eugene, OR) (3). The amount of RNA was quantified by absorption at 260 nm (52).…”

Section: Rna Isolation and Rt-pcrmentioning

confidence: 99%

Prostaglandin E₂-synthesizing enzymes in fever: differential transcriptional regulation

Ivanov¹,

Pero

Scheck

et al. 2002

American Journal of Physiology-Regulatory, Integrative and Comp

131

116

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The febrile response to lipopolysaccharide (LPS) consists of three phases (phases I-III), all requiring de novo synthesis of prostaglandin (PG) E(2). The major mechanism for activation of PGE(2)-synthesizing enzymes is transcriptional upregulation. The triphasic febrile response of Wistar-Kyoto rats to intravenous LPS (50 microg/kg) was studied. Using real-time RT-PCR, the expression of seven PGE(2)-synthesizing enzymes in the LPS-processing organs (liver and lungs) and the brain "febrigenic center" (hypothalamus) was quantified. Phase I involved transcriptional upregulation of the functionally coupled cyclooxygenase (COX)-2 and microsomal (m) PGE synthase (PGES) in the liver and lungs. Phase II entailed robust upregulation of all enzymes of the major inflammatory pathway, i.e., secretory (s) phospholipase (PL) A(2)-IIA --> COX-2 --> mPGES, in both the periphery and brain. Phase III was accompanied by the induction of cytosolic (c) PLA(2)-alpha in the hypothalamus, further upregulation of sPLA(2)-IIA and mPGES in the hypothalamus and liver, and a decrease in the expression of COX-1 and COX-2 in all tissues studied. Neither sPLA(2)-V nor cPGES was induced by LPS. The high magnitude of upregulation of mPGES and sPLA(2)-IIA (1,257-fold and 133-fold, respectively) makes these enzymes attractive targets for anti-inflammatory therapy.

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Section: Rna Isolation and Rt-pcrmentioning

confidence: 99%

Prostaglandin E₂-synthesizing enzymes in fever: differential transcriptional regulation

Ivanov¹,

Pero

Scheck

et al. 2002

American Journal of Physiology-Regulatory, Integrative and Comp

131

116

View full text Add to dashboard Cite

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“…The visualisation results of agarose gel electrophoresis conducted in this study showed three bands (Supplementary data 10). According to Rapley and Heptinstall (1998), two bands of the three bands may show ribosomal RNA of 18S rRNA and 28S rRNA, which are the largest components in total RNA. The thickness (intensity) and sharpness of the tape fluorescence could show the quality of the total RNA isolate well enough.…”

Section: Discussionmentioning

confidence: 99%

Expression of gag-CA Gene of Jembrana Disease Virus with Cationic Liposomal and Chitosan Nanoparticle Delivery Systems as DNA Vaccine Candidates

Unsunnidhal¹,

Ishak²,

Kusumawati³

2019

TLSR

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Highlights • The gag-CA gene from the Jembrana Disease Virus was successfully optimised and constructed in the pEGFP-C1 vector to produce recombinant DNA pEGFP-C1-gag-CA. • Chitosan successfully encapsulates the recombinant DNA producing nanoparticle using mass ratio of DNA with chitosan 1:2 to 1:4. • The recombinant DNA vaccine was successfully delivered and expressed in cell line with cationic liposomal and chitosan nanoparticle delivery systems.

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“…Kuantitas DNA dalam suatu sampel dapat dihitung berdasarkan nilai absorbansi pada panjang gelombang tersebut. Sebuah sampel memiliki nilai absorbansi 1 pada λ260 nm memiliki konsentrasi DNA sebanyak 50µg/mL (Rapley and Heptinstall, 1998 Amplifikasi gen 28S rDNA memiliki kualitas pita produk yang terlihat terang dan tebal, serta sejajar satu sama lain dimana menghasilkan pola pita tunggal DNA yang menandakan bahwa primer dan suhu annealing yang digunakan sudah tepat. Hasil amplifikasi PCR fragmen gen pengkode 28S rDNA menggunakan pasangan primer ITS1 dan ITS4 seperti pada Gambar 4.…”

Section: Konsentrasi Dna C Tropicalis Menggunakan Spektrofotometri Uunclassified

AMPLIFIKASI PCR DOMAIN D1/D2 28S rDNA MENGGUNAKAN PRIMER ITS1 DAN ITS4 SAMPEL DNA DARI Candida tropicalis YANG DIISOLASI DENGAN METODE PENDINGINAN

Hermansyah

Sutami

Miksusanti

2018

IJoPAC

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The purpose of this research was to isolated DNA from the yeast C. tropicalis with freeze thawing method -200 C conducted on 3 colonies of C. tropicalis. Each colony threated variations of cooling, 3x15 minutes, 3x25 minutes and 3x35 minutes, to break the cell walls. Subsequently all the samples amplified with 3 variations of PCR cycles, 15 cycles, 25 cycles and 35 cycles, after all of the samples isolated by freeze thawing method -200 C. Its was known that sample A15 has the smallest concentration of DNA yeast C. tropicalis, ie 50 µg/mL, while sample C35 had the largest concentration of DNA yeast C. tropicalis, ie 225 µg/mL. The result of the research indicated that the best condition can be reached in 3x35 minutes. On 35th cycle has clearer C. tropicalis DNA bands than the 25th and 15th PCR cycle. C. tropicalis DNA bands at 35th cycles there were 7 DNA bands were detected and bright bands on a long 35 minutes cooling. In the 25th and the 15th cycle, there was no DNA bands were detected in all samples. Based on the results obtained, the amplification process must be carried out at least 35 times cycles so that the C. tropicalis DNA bands can be detected.

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UV Spectrophotometric Analysis of Ribonucleic Acids

Cited by 6 publications

References 0 publications

Prostaglandin E₂-synthesizing enzymes in fever: differential transcriptional regulation

Prostaglandin E₂-synthesizing enzymes in fever: differential transcriptional regulation

Expression of gag-CA Gene of Jembrana Disease Virus with Cationic Liposomal and Chitosan Nanoparticle Delivery Systems as DNA Vaccine Candidates

AMPLIFIKASI PCR DOMAIN D1/D2 28S rDNA MENGGUNAKAN PRIMER ITS1 DAN ITS4 SAMPEL DNA DARI Candida tropicalis YANG DIISOLASI DENGAN METODE PENDINGINAN

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UV Spectrophotometric Analysis of Ribonucleic Acids

Cited by 6 publications

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Prostaglandin E2-synthesizing enzymes in fever: differential transcriptional regulation

Prostaglandin E2-synthesizing enzymes in fever: differential transcriptional regulation

Expression of gag-CA Gene of Jembrana Disease Virus with Cationic Liposomal and Chitosan Nanoparticle Delivery Systems as DNA Vaccine Candidates

AMPLIFIKASI PCR DOMAIN D1/D2 28S rDNA MENGGUNAKAN PRIMER ITS1 DAN ITS4 SAMPEL DNA DARI Candida tropicalis YANG DIISOLASI DENGAN METODE PENDINGINAN

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Prostaglandin E₂-synthesizing enzymes in fever: differential transcriptional regulation

Prostaglandin E₂-synthesizing enzymes in fever: differential transcriptional regulation