• Title/Summary/Keyword: Nucleic acid extraction

Search Result 25, Processing Time 0.022 seconds

Analysis of Cellular Components of Starch-Utilizing Yeast Sporobolomyces holsaticus (전분이용성 효모 Sporobolomyces holsaticus의 균체성분 분석에 대하여)

  • Park, Wan-Soo;Koo, Young-Jo;Shin, Dong-Hwa;Suh, Kee-Bong
    • Korean Journal of Food Science and Technology
    • /
    • v.15 no.2
    • /
    • pp.170-176
    • /
    • 1983
  • Starchy single cell protein produced by a starch-utilizing yeast, Sporobolomyces holsaticus FRI Y-5 was analyzed for its composition such as intracellular protein, amino acids, fatty acids, minerals, vitamins and pigments. It was shown that it contained 33.08% of total carbohydrate, 45.63% of crude protein, 20.01% of crude lipid, 3.24% of ash and 4.46% of pigment. Whole cell extracted by cold and hot NaOH method contained 40.89% of soluble protein and the estimated nucleic acid content from crude and soluble protein contents was about 7.6%. The sulphur-containing amino acids, threonine, isoleucine and valine were analyzed to be the limiting amino acids in the starchy SCP, and the protein score was calculated as 89.4. It was shown from its fatty acid analysis that it contained $6.5%\;of\;C_{16:0}$, $2.4%\;of\;C_{18:0}$, $81.9%\;of\;C_{18:1}$, $3.2%\;of\;C_{18:2}$, and $6.0%\;of\;C_{18:3}$. Also it was observed that it contained, per 100 g of dry cell, 365.33mg of Mg and 282.75mg of K more than Fe and Ca. The content of Vit. $B_2$ was 3.7mg per 100 g of dry cell, but niacin was not detected under this experimental condition. The UV-visible scanning result of pigment extract showed that the yeast contained carotenoid and unknown pigments.

  • PDF

Fractionation of the Cells of Staphylococcus aureus (Staphylococcus aureus의 균체분획(菌體分劃))

  • Choi, Kyoung-Ho;Hyun, Eun-Min;Park, Kum-Soon
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.10 no.1
    • /
    • pp.85-91
    • /
    • 1981
  • The growing cells of S. aureus were fractionated along the Schmidt-Thannhauser-Schneider's technique into several fractions such as TCA(trichloroacetic acid)-soluble, lipid, nucleic acid, protein and residue fraction. They were also fractionated according to their cellular structure into Sonic-supernatant, SDS(sodium lauryl sulfate)-soluble, Formamide-soluble and Residue fraction. Fractionation was carried out by orderly treatment of the Sonic pellet with 1.0% SDS and hot$(150^{\circ}C)$ formamide, and the pellet was prepared by centrifugation of the cells sonic osillated for 20 minutes at 150 watt. Sonic-supernatant fraction contained a 91.3% of total DNA while other fractions contained less than 9.5%. SDS-soluble fraction showed a high activity of malate dehydrogenase(13.67 unit/mg protein) and which was higher 22.3 times than the activity found from unsoluble fraction. Formamide-soluble fraction prepared from SDS-undoluble pellet by using the hot formamide exhibited a clear action of reducing sugars against the Anthronesulfate, while it exhibited no clear action against the ninhydrin. However, contrastly, the residue remainnning after extraction with formamide exhibited a clear action against ninhydrin and glucosamine was detected form the hydrolysate of residue by paper chromatography. From these results it is considered that the Sonic-supernatant fraction is mainly consisted of plasmic component of the cells. Other fractions, SDS-soluble, Formamide-soluble and Residue, should be consisted of plasma membrane, lipoplysaccharide and peptidoglycan of the cell, respectively.

  • PDF

Convenient Virion Capture (VC)/PCR for Tomato yellow leaf curl geminivirus Occurring on Tomato in Korea (우리나라 토마토에 발생한 토마토황화잎말림바이러스(Tomato yellow leaf curl geminivirus)의 초간편 Virion Capture(VC)/PCR 진단법)

  • Cho, Jeom-Deog;Kim, Tae-Seong;Kim, Ju-Hee;Choi, Gug-Seoun;Chung, Bong-Nam;Choi, Hong-Soo;Kim, Jeong-Soo
    • Research in Plant Disease
    • /
    • v.14 no.3
    • /
    • pp.233-237
    • /
    • 2008
  • Tomato yellow leaf curl virus (TYLCV), a newly reported Geminivirus from tomato, generated recently large economic losses in Korea. Development of a fast and precise genetic diagnosis technique for detecting TYLCV which Agricultural research and extension services can utilize easy and handy is very important to prevent yield losses. Virion Capture (VC)/PCR is a simple, accurate and economical genetic detection method without any works or commercial kits for the extraction of the nucleic acid from the infected plants. Primers of twenty two for detection of TYLCV were designed and tested with extracted total DNA or crude sap from tomato leaf infected with TYLCV and healthy plant. Nine primers for total DNA using conventional PCR and another 9 primers for VC/PCR were selected eventually. Primers of six having same specificity were selected from the two methods and tested with other Geminivirus, Tobacco leaf curl virus (TLCV) by VC/PCR. Finally specific primers of four were selected for detection of TYLCV using VC/PCR, and Deng (540, 541), a degenerate primer for Geminivirus reported in 1996, was also developed for VC/PCR.

Development of HLA-A, -B and -DR Typing Method Using Next-Generation Sequencing (차세대염기서열분석법을 이용한 HLA-A, -B 그리고 -DR 형별 분석법 개발)

  • Seo, Dong Hee;Lee, Jeong Min;Park, Mi Ok;Lee, Hyun Ju;Moon, Seo Yoon;Oh, Mijin;Kim, So Young;Lee, Sang-Heon;Hyeong, Ki-Eun;Hu, Hae-Jin;Cho, Dae-Yeon
    • The Korean Journal of Blood Transfusion
    • /
    • v.29 no.3
    • /
    • pp.310-319
    • /
    • 2018
  • Background: Research on next-generation sequencing (NGS)-based HLA typing is active. To resolve the phase ambiguity and long turn-around-time of conventional high resolution HLA typing, this study developed a NGS-based high resolution HLA typing method that can handle large-scale samples within an efficient testing time. Methods: For HLA NGS, the condition of nucleic acid extraction, library construction, PCR mechanism, and HLA typing with bioinformatics were developed. To confirm the accuracy of the NGS-based HLA typing method, the results of 192 samples HLA typed by SSOP and 28 samples typed by SBT compared to NGS-based HLA-A, -B and -DR typing. Results: DNA library construction through two-step PCR, NGS sequencing with MiSeq (Illumina Inc., San Diego, USA), and the data analysis platform were established. NGS-based HLA typing results were compatible with known HLA types from 220 blood samples. Conclusion: The NSG-based HLA typing method could handle large volume samples with high-throughput. Therefore, it would be useful for HLA typing of bone marrow donation volunteers.

A Study on STR Analysis According to the Method of Developing Latent Fngerprints Deposited on Non-Porous Surfaces in the Marine Environment (해양환경 내 비다공성 표면에 유류된 잠재지문 현출방법에 따른 STR 분석 연구)

  • Kim, Jin-Sun;Kim, Sea-In;Yoon, Hyun-Kyoung;Choo, Min-kyu
    • The Journal of the Korea Contents Association
    • /
    • v.22 no.10
    • /
    • pp.733-741
    • /
    • 2022
  • Among the various evidence found in maritime crimes, fingerprints and DNA are very important in that they can identify a suspect. In this study, 5 types of non-porous surfaces (plastic, stainless, glass, ceramic, FRP), which are often found as evidence in the actual marine environment, were selected, and latent and blood fingerprints were passed down and immersed at the Donghae Maritime Police Station's exclusive pier for about 7 days. After that, DNA extraction, quantification, and STR profile were analyzed after fingerprint developing CA fumming method and 4 powder methods (Swedish black powder, Concentrated black powder, Supranano red powder, Dazzle orange powder). Among the fingerprint developing methods, when Supranano red powder was applied, a relatively high amount of DNA was found. As a result of STR profile analysis, an average of 16.8 to 9 loci were secured, and all 20 were confirmed in glass and ceramic materials. As a result of the study, it was possible to secure the STR profile by extracting and quantifying DNA after applying the fingerprint developing method to virtual evidence immersed for about 7 days, and further research is needed to secure the STR profile by analyzing DNA after applying various fingerprint developing methods such as VMD and SPR.