Spin-Column Chromatography for DNA Purification

1996 ◽  
Vol 239 (1) ◽  
pp. 117-118 ◽  
Author(s):  
Nels E. Pederson
Keyword(s):  
Column Chromatography ◽  
Dna Purification ◽  
Spin Column

Sepharose spin column chromatography

1994 ◽  
Vol 1 (1) ◽  
pp. 105-108 ◽  
Author(s):  
Jac A. Nickoloff
Keyword(s):  
Column Chromatography ◽  
Spin Column

scholarly journals Simple, inexpensive and RNase-free purification of plasmid DNA by fractional precipitation with isopropanol

BioTechniques ◽  
2021 ◽  
Author(s):  
Viiu Paalme ◽  
Mart Speek
Keyword(s):  
Quality Control ◽  
Column Chromatography ◽  
Plasmid Dna ◽  
Fractional Precipitation ◽  
Alkaline Lysis ◽  
Bacterial Extract ◽  
Total Rna ◽  
Spin Column ◽  
Commercial Kits

We present a modified alkaline lysis method for purification of plasmid DNA (pDNA) from bacterial extract using fractional precipitation with isopropanol (FPI). This method includes two successive precipitations with 0.33 and 0.36 volumes of isopropanol and separates pDNA from total RNA and most of the lipopolysaccharides. Using different quality control tests, we demonstrate that plasmids purified with FPI show superior quality compared to plasmids prepared with commercial kits based on spin-column chromatography.

Solution-Based Fabrication of Carbon Nanotube Gas Sensor by Using Dielectrophoresis and Spin-Column Chromatography

2013 ◽  
Vol 699 ◽  
pp. 915-920
Author(s):  
Hideaki Watanabe ◽  
Hiroki Komure ◽  
Michihiko Nakano ◽  
Junya Suehiro
Keyword(s):  
Carbon Nanotubes ◽  
Electronic Structure ◽  
Carbon Nanotube ◽  
Gas Sensor ◽  
Column Chromatography ◽  
Gas Sensors ◽  
Single Walled Carbon Nanotubes ◽  
Sensor Response ◽  
Spin Column ◽  
Walled Carbon Nanotubes

Single-walled carbon nanotubes (SWCNTs) gas sensor has attracted a great deal of attention because of their remarkable properties. The sensor response is attribute to the semiconducting CNT whose electronic properties depend on its chirality. The authors have previously found that the sensor response increased by using separated semiconducting SWCNTs from a mixture with metallic one. Since the electronic structure (metallic or semiconducting) of CNTs is governed by their chirality, a chirality-selective fabrication of CNT gas sensor is essential to improve their performance. In this study, we proposed chirality-based separation of semiconducting SWCNTs by using spin-column chromatography. Pristine CNT suspension was separated into three fractions that had different chiralities of semiconducting SWCNTs. Separated semiconducting CNTs of each fraction were used for fabrication of three CNT gas sensors by dielectrophoresis. Comparison of these sensor responses to NO2 revealed that sensor response depended on the chirality.

Dielectrophoretic Assembly of Semiconducting Carbon Nanotubes Separated and Enriched by Spin Column Chromatography and Its Application to Gas Sensing

2012 ◽  
Vol 51 ◽  
pp. 045102 ◽  
Author(s):  
Michihiko Nakano ◽  
Masahiro Fujioka ◽  
Kaori Mai ◽  
Hideaki Watanabe ◽  
Yul Martin ◽  
...  
Keyword(s):  
Carbon Nanotubes ◽  
Column Chromatography ◽  
Gas Sensing ◽  
Spin Column

Dielectrophoretic Assembly of Semiconducting Carbon Nanotubes Separated and Enriched by Spin Column Chromatography and Its Application to Gas Sensing

2012 ◽  
Vol 51 (4R) ◽  
pp. 045102 ◽  
Author(s):  
Michihiko Nakano ◽  
Masahiro Fujioka ◽  
Kaori Mai ◽  
Hideaki Watanabe ◽  
Yul Martin ◽  
...  
Keyword(s):  
Carbon Nanotubes ◽  
Column Chromatography ◽  
Gas Sensing ◽  
Spin Column

scholarly journals Comparison of DNA extraction methods for molecular identification of pathogenic Leptospira in the urine samples

2020 ◽  
Vol 11 (2) ◽  
pp. 77-84
Author(s):  
Farida Dwi Handayani ◽  
Rahmi Ayu Wijayaningsih ◽  
Muhammad Hussein Gasem ◽  
Tri Wibawa
Keyword(s):  
Dna Extraction ◽  
Urine Sample ◽  
Column Chromatography ◽  
Dna Isolation ◽  
Storage Temperature ◽  
Dna Amplification ◽  
Life Time ◽  
Pathogenic Leptospira ◽  
Guanidine Isothiocyanate ◽  
Spin Column

Latar belakang: Leptospirosis merupakan zoonosis penting di dunia, yang masih sering terjadi salah diagnosis. Deteksi laboratorium Leptospira menjadi tantangan karena bakterimea cukup singkat untuk dideteksi molekuler, namun antibodi juga muncul sangat lambat. Urine dapat menjadi sampel alternatif untuk deteksi PCR pada leptospirosis. Pengerjaan PCR membutuhkan DNA berkualitas dan andal, dan diperoleh dari metode ekstraksi DNA yang baik. Penelitian bertujuan untuk mengetahui metode ekstraksi DNA Leptospira terbaik untuk sampel urin, serta mengevaluasi pengaruh waktu penyimpanan dan suhu terhadap kestabilan DNA. Metode: Penelitian ini menggunakan tiga metode isolasi DNA yang berbeda; berbasis silika dengan spin kolom, kromatografi spin column menggunakan resin sebagai matriks pemisah, dan metode larutan dengan guanidine isothiocyanate. Hasil ekstraksi diperiksa konsentrasi dan kemurniannya. Gen SecY pada Leptospira dideteksi dengan PCR real-time. Pengaruh suhu dan lama penyimpanan DNA juga dilihat. Hasil: Hasil isolasi DNA menggunakan resin menunjukkan konsentrasi tertinggi (7,94 + 2,11 μg / mL) dan jumlah salinan amplifikasi DNA Leptospira tertinggi (50167,92 + 1,19). Suhu penyimpanan pada suhu 4°C, -20°C, dan -80°C dan umur simpan 91 hari tidak berpengaruh terhadap kualitas dan kuantitas DNA Leptospira hasil isolasi spike urin. Kesimpulan: Isolasi DNA menggunakan spin column chromatography dengan resin sebagai matriks separasi memiliki kualitas dan kuantitas terbaik berdasarkan kemurnian dan konsentrasi DNA serta jumlah gen SecY yang teramplifikasi. Kata kunci: Leptospira, Leptospirosis, ekstraksi DNA, sampel urin, penyimpanan sampel.   Abstract Background: Leptospirosis is a worldwide zoonotic disease, which is still often misdiagnosed. Laboratory detection of Leptospira is challenging since the bacteraemia is quite short for molecular detection, however, the rise of the antibody is late to post the infection. Urine can be a potential alternative sample for PCR detection in leptospirosis. The PCR method requires a reliable DNA template, which is obtained from good DNA extracting methods. The study aimed to determine the best method of extraction Leptospira DNA from the urine sample, as well as evaluating the effect of time storage and temperature for its DNA stability. Methods: This study was utilizing three different DNA isolation methods; silica based with spin column, spin column chromatography using resin as separation matrix, and solution method with guanidine isothiocyanate. The yields were examined for its concentration and purity. Leptospira’s SecY gene was detected with realtime PCR. The influences of storage temperature and the life time of the DNA were also studied. Results: The yield of DNA isolation using resin showed the highest concentration (7.94+2.11 μg/mL) and highest Leptospira DNA amplification copy number (50167.92+1.19). Storage temperature at 4°C, -20°C, and -80°C and life time of 91 days did not have any effect on the quality and quatnity of Leptospira DNA isolated from spiked urine. Conclusions: DNA isolation using spin column chromatography with resin as separation matrix has the best quality and quantity based on the purity and concentration of DNA and the higher number of amplified SecY gene. Keywords: Leptospira, Leptospirosis, DNA extraction, urine sample, sample storage  

scholarly journals Polyphenol oxidase from wheat bran is a serpin.

2008 ◽  
Vol 55 (2) ◽  
pp. 325-328 ◽  
Author(s):  
Yoshiki Yamasaki ◽  
Haruyoshi Konno ◽  
Kazuhiko Noda
Keyword(s):  
Polyphenol Oxidase ◽  
Column Chromatography ◽  
Wheat Bran ◽  
Serine Proteases ◽  
Batch Adsorption ◽  
Amino Acid Residues ◽  
Protein Z ◽  
Spin Column ◽  
Sds Page ◽  
Relative Molecular Weight

Polyphenol oxidase (PPO; EC 1.10.3.2) was isolated from wheat bran by a procedure that included ammonium sulfate fractionation, batch adsorption by DEAE-cellulofine, CM-cellulofine column chromatography, DEAE-cellulofine column chromatography, preparative isoelectric focusing, adsorption on the membrane of a Vivapure Q Maxi H spin column, and heat treatment. These procedures led to 150-fold purification with 4.2% recovery. The PPO was homogeneous by SDS/PAGE. The relative molecular weight of the PPO was estimated to be 37,000 based on its mobility in SDS/PAGE. The isoelectric point of the PPO was 4.4. The K(m) values of the PPO for caffeic acid, chlorogenic acid, pyrocatechol, 4-methyl catechol and l-DOPA as substrates were 0.077, 0.198, 1.176, 1.667 and 4.545 mM. The PPO was strongly inhibited by tropolone. The K(i) value for tropolone is 2.2 x 10(-7) M. The sequence of the 15 N-terminal amino-acid residues was determined to be ATDVRLSIAHQTRFA, which was identical to those of serpin from Triticum aestivum and protein Z from Hordeum vulgare. The PPO strongly inhibited the activity of trypsin, which is an enzyme of serine proteases; 50% inhibition was observed with 1.5 x 10(-7) M PPO. The K(i) value for PPO is 2.3 x 10(-8) M. The wheat bran PPO should be a very important protein for protecting wheat against disease, virus, insect and herbivore damages by both the activities of PPO and protease inhibitor.

Heparin Excretion in Intact and Hepatectomized Rats

1981 ◽  
Vol 45 (02) ◽  
pp. 146-149
Author(s):  
Ray Losito ◽  
Harry Gattiker ◽  
Ginette Bilodeau
Keyword(s):  
Column Chromatography ◽  
Normal Values ◽  
Anticoagulant Activity ◽  
Biliary Fistula ◽  
Kinetics Of ◽  
Intact Rats

SummaryMetabolism and kinetics of 3H-heparin were compared in intact and hepatectomized rats. Rats were divided into three groups: 1) intact rats with biliary fistulas and cystostomies 2) intact rats with only cystostomies and 3) hepatectomized rats with cystostomies. Radioactivity in blood, bile and urine besides anticoagulant activity in blood and urine were examined. In addition, column chromatography of urine was used to isolate possible metabolites. Seventy percent and 80% of the radioactive dose was found in the urine of intact rats at 24 hr and 48 hr. Close to 5% of the radioactivity was found in bile or rats with a biliary fistula after 48 hr. The APTT declined to near normal values at 1 hr whether rats had a biliary fistula or not. In contrast, only 25 % of the radioactivity could be excreted into the urine of hepatectomized rats in 24 hr; the APTT did not decline as fast and at 5 hr, it was still 100 seconds. Only one radioactive component could be isolated on chromatography from all urines of these animals and appears to be similar to the original heparin. Thus, the liver has an important role to play in regulating the anticoagulant effects and excretion of heparin.

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