• Journal of Plant Biology
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• v.30 no.3
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• pp.201-203
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• 1987

Total RNA was isolated from two months old wheat, rice, tobacco and sweet potato. The procedure used was simple and provided pure RNA preparation. Lysis of plant tissue in a buffer with guanidine thiocyanate and CsCl density gradient centrifugation separated RNA from the rest of the cellular components. Subsequent cholroform/1-butanol extraction and ethanol precipitation were necessary to ensure contaminant-free RNA preparation. Storage of the lysed plant tissue in the buffer with guanidine thiocyanate preserved the sample for two months without noticeable RNA degradation.

• Molecular & Cellular Toxicology
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• v.1 no.2
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• pp.92-98
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• 2005

DNA microarray analysis of gene expression in toxicogenomics typically requires relatively large amounts of total RNA. This limits the use of DNA microarray when the sample available is small. To confront this limitation, different methods of linear RNA amplification that generate antisense RNA (aRNA) have been optimized for microarray use. The target preparation strategy using amplified RNA in DNA microarray protocol can be divided into direct-incorporation labeling which resulted in cDNA targets (Cy-dye labeled cDNA from aRNA) and indirect-labeling which resulted in cRNA targets (i.e. Cy-dye labeled aRNA), respectively. However, despite the common use of amplified targets (cDNA or cRNA) from aRNAs, no systemic assessment for the use of amplified targets and bias in terms of hybridization performance has been reported. In this investigation, we have compared the hybridization performance of cRNA targets with cDNA targets from aRNA on a 10 K cDNA microarrays. Under optimized hybridization conditions, we found that 43% of outliers from cDNA technique and 86% from the outlier genes were reproducibly detected by both targets hybridization onto cDNA microarray. This suggests that the cRNA labeling method may have a reduced capacity for detecting the differential gene expression when compared to the cDNA target preparation. However, further validation of this discordant result should be pursued to determine which techniques possesses better accuracy in identifying truly differential genes.

• BMB Reports
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• v.37 no.3
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• pp.351-355
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• 2004

Separate protocols are commonly used to prepare plasmid DNA, chromosomal DNA, or total RNA from E. coli cells. Various methods for the rapid preparation of plasmid DNA have been developed previously, but the preparation of the chromosomal DNA and total RNA are usually laborious. We report here a simple, fast, reliable, and cost-effective method to extract total nucleic acids from E. coli by direct lysis of the cells with phenol. Five distinct and sharp bands, which correspond to chromosomal DNA, plasmid DNA, 23S rRNA, 16S rRNA, and a mixture of small RNA, were observed when analyzing the prepared total nucleic acids on a regular 1-2% agarose gel. The simple and high-quality preparation of the total nucleic acids in a singe tube allowed us to rapidly screen the recombinant plasmid, as well as to simultaneously monitor the change of the plasmid copy number and rRNA levels during the growth of E. coli in the liquid medium.

• KSBB Journal
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• v.17 no.3
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• pp.311-313
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• 2002

Lactobacillus spry. are Gram-positive bacteria playing important roles in human health. In this study, we successfully isolated the total RNA from the cells broken by glass beads using hot phenol method. Moreover, we were able to omit lysozyme and proteinase K treatment by using glass beads to break cell more efficiently. This method was more rapid and simple when compared to the previous one. Prepared RNA can be used for the transcriptional analysis of Lactobacillus spp.

• Journal of the Korean Chemical Society
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• v.50 no.3
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• pp.269-271
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• 2006

• Biomedical Science Letters
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• v.2 no.2
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• pp.257-265
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• 1996

Simple stain and electron microscopic in situ hybridization is studied and applied for the identification of rabbit haemorrhagic disease viral RNA in a unicrylated preparation of the liver after innoculation of rabbit haemorrhagic disease virus. Hybridization for detection of viral RNA in unicryl embedded tissues using complementary 84 bases oligonucleotide probe labelled by biotin CE-phosphoramidite compared with 4717∼4800 sequences of rabbit haemorrhagic disease virus, modified hybridization protocol and antibiotin antibody-l0nm gold as signal marker. The best results were obtained in 0.02% glutaraldehyde, Unicryl resin cell block, biotinylated oligonucleotide probes, antibiotin-l0nm gold. In this report, RHD viral RNA was distributed widely within the mitochondria and nucleus of liver cell by electron microscopic in situ hybridization. In situ hybridization has become a standard method for localizing DNA or RNA sequences in tissue or celt preparation. In situ hybridization is detected the virus genome in the cells and tissue as specifically compared with others nucleic acid hybridization method.

• Bulletin of the Korean Chemical Society
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• v.33 no.11
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• pp.3861-3863
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• 2012

• Journal of Life Science
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• v.16 no.2 s.75
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• pp.240-244
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• 2006

We present a fast and simple protocol for purification of mitochondria, mitochondrial DNA, and RNA from small amounts of tomato leaves. This method uses a high ionic strength medium to isolate mitochondria and extract mitochondrial DNA and RNA from a single preparation and is easily adaptable to other plant species. Mitochondria was confirmed by MitoTracker. The mitochondrial DNA was not contaminated by plastid DNA, was successfully used for PCR. Similarly, the isolated mitochondrial RNA was not contaminated only slightly contaminated (leaves) by plastid RNA. RNA prepared according to our method was acceptable for RT-PCR analysis

• Journal of the Korea Convergence Society
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• v.12 no.8
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• pp.179-186
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• 2021

Among the Covid-19 vaccine platforms, mRNA-platform vaccines are summarized qualitatively in this paper. Manufacturing mRNA vaccines consist of serial processes; the preparation process of DNA template, the transcription of mRNA, nanoemulsion process, and the fill and finish unit combined with formulation stages. It is noticeable that major players are collaborated for producing mRNA vaccines. In particular, the nanoemulsion process is recognized to the key process requiring formulated lipid materials to protect modified mRNA until they arrive in intracellular cytosol. It is known that the nanoemulsion process adapts well-designed microfluidic devices. We expect that the nanoemulsion process will stimulate pharmaceutical industries to develop diverse applications.

• Microbiology and Biotechnology Letters
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• v.25 no.5
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• pp.512-519
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• 1997

RNA accumulating strain of Torulopsis versatilis MT-1 was cultured in molasses medium for higher contents of RNA in cell. Yeast cells were harvested at logarithmic phase on synchronous culture. Yield of cells on dry base to input sugar was 59.5%. Crude protein content was 55.1% in cell. RNA content was 13.9%. Some problems found in the process for the preparation of yeast extracts were improved by the addition of culture broth of Streptomyces faecalis MSF which secrete RNase (5' nuclease and 5' adenylic acid deaminase). When the culture broth of S. faecalis MSF was added in autolysis process 46% of RNA in cell was converted to I and G(5' inosinic acid and 5' guanylic acid) in extract. By addition of 3-7% culture broth of S.faecalis MSF in autolysis or enzymolysis process at the start or early stage, RNA in extract was converted easily to I and G and protein in cells was easily extracted and hydrolyzed to amino acid. Taste of those yeast extracts was delicious. The yeasty smell in yeast extracts was removed. And cell debris was easily removed from extract.