Affinity Separation of Nucleic Acids on Monosized Magnetic Beads

1993 ◽  
pp. 479-485
Author(s):  
Mathias Uhlén ◽  
Ørjan Olsvik ◽  
Erik Hornes
Keyword(s):  
Nucleic Acids ◽  
Magnetic Beads ◽  
Affinity Separation

scholarly journals Adsorption and isolation of nucleic acids on cellulose magnetic beads using a three-dimensional printed microfluidic chip

2015 ◽  
Vol 9 (6) ◽  
pp. 064118 ◽  
Author(s):  
Lei Zhang ◽  
Rachel N. Deraney ◽  
Anubhav Tripathi
Keyword(s):  
Nucleic Acids ◽  
Microfluidic Chip ◽  
Magnetic Beads ◽  
Three Dimensional

High Throughput RNA Extraction and PCR Inhibitor Removal of Settled Solids for Wastewater Surveillance of SARS-CoV-2 RNA v2

2021 ◽  
Author(s):  
Aaron Topol ◽  
marlene.wolfe not provided ◽  
Krista Wigginton ◽  
Bradley White ◽  
Alexandria B Boehm
Keyword(s):  
Nucleic Acids ◽  
High Throughput ◽  
Magnetic Beads ◽  
Rna Extraction ◽  
Poly Vinyl Alcohol ◽  
Enzymatic Reactions ◽  
Efficient Removal ◽  
Dna And Rna ◽  
Pcr Inhibitor ◽  
Wastewater Samples

Please note that while this protocol is for TNA extraction using the Perkin Elmer Chemagic 360, RNA extraction with resuspended solids from this protocol has been verified to perform well using the Kingfisher MagMax kit as another high throughput, automated option and two manual Qiagen kits - the All Prep Powerviral DNA/RNA Kit and the Qiamp Viral RNA Mini Kit. This process instruction describes the steps for purification of nucleic acids from wastewater solids and preparation for downstream quantitative analysis with Reverse Transcriptase droplet digital Polymerase Chain Reaction (RT-ddPCR). Due to the large quantities of substances that have inhibitory effects on PCR in wastewater samples, a subsequent PCR inhibitor removal step is required after nucleic acid purification. Both steps of the process are carried out in a 96-well plate format. This method uses the resuspended solids generated using this protocol: High Throughput pre-analytical processing of wastewater settled solids for SARS-CoV-2 RNA analyses. RNA purification is carried out using a kit optimized for the purification of viral on for the Perkin Elmer Chemagic 360. Although only RNA is used in the downstream applications from this protocol, DNA is also eluted in this process. A crucial component of the purification kit are the magnetic particles coated with poly vinyl alcohol (M-PVA Magnetic Beads) which have a hydrophilic surface giving them an affinity for nucleic acids but not many other biological molecules. The workflow involves binding nucleic acids in a sample to the beads which are then transferred through a series of wash buffers to remove debris with a robotic head with magnetic rods. The OneStep PCR Inhibitor Removal Kits are PCR inhibitor clean up kits that contain all the components needed for efficient removal of contaminants that can inhibit downstream enzymatic reactions (e.g. PCR and RT) from DNA and RNA preparations. The column matrices in these PCR inhibitor clean up kits have been specifically designed for the efficient removal of polyphenolic compounds, humic/fulvic acids, tannins, melanin, etc. from the most impure DNA and RNA preparations. This process instruction applies to extraction of RNA from wastewater samples using the Chemagic™ Viral DNA/RNA 300 Kit H96 for the Perkin Elmer Chemagic 360 followed by PCR Inhibitor Removal with the Zymo OneStep-96 PCR Inhibitor Removal Kit.

Ni2+-zeolite/ferrosphere and Ni2+-silica/ferrosphere beads for magnetic affinity separation of histidine-tagged proteins

2016 ◽  
Vol 45 (4) ◽  
pp. 1582-1592 ◽  
Author(s):  
T. A. Vereshchagina ◽  
M. A. Fedorchak ◽  
O. M. Sharonova ◽  
E. V. Fomenko ◽  
N. N. Shishkina ◽  
...  
Keyword(s):  
Recombinant Proteins ◽  
Shell Structure ◽  
Magnetic Beads ◽  
Core Shell ◽  
Affinity Separation ◽  
High Affinity ◽  
Core Shell Structure

Ferrosphere-based magnetic beads of a core–shell structure with immobilized Ni2+ having a high affinity towards His-tagged recombinant proteins were fabricated.

scholarly journals The trans-Acting Protein Interacting with the DNA Motif Proximal to the Transcriptional Start Site of Plant l-Asparaginase Is Bacterial Sarcosine Oxidase

2004 ◽  
Vol 186 (3) ◽  
pp. 811-817 ◽  
Author(s):  
William T. Jones ◽  
Taha Al-Samarrai ◽  
Janice M. Reeves ◽  
Gordon B. Ryan ◽  
Christopher A. Kirk ◽  
...  
Keyword(s):  
Magnetic Beads ◽  
Transcriptional Start Site ◽  
Sequence Motif ◽  
Start Site ◽  
Affinity Separation ◽  
Specific Sequence ◽  
Repressor Protein ◽  
Α Subunit ◽  
Sarcosine Oxidase ◽  
Gel Retardation

ABSTRACT A trans-acting protein interacting with a specific sequence motif proximal to the transcriptional start site of the l-asparaginase promoter has been observed previously (E. Vincze, J. M. Reeves, E. Lamping, K. J. F. Farnden, and P. H. S. Reynolds, Plant Mol. Biol. 26:303-311, 1994). Gel retardation experiments in which protein extracts of Mesorhizobium loti and developing nodules were used suggested a bacterial origin for the repressor binding protein (rep2037). Nodulation tests were performed by using different Fix− Tn5 mutants of M. loti. Analyses of these mutants revealed a correlation between the presence of Mesorhizobium in the nodule-like structures and the ability of nodule protein extracts to bind the repressor binding domain (RBD). Through the use of mutated RBD sequences, the RBD sequence was identified as CTAAAAT. The repressor protein was isolated from M. loti NZP2037 by multiple chromatographic procedures and affinity separation by using concatemers of RBD attached to magnetic beads. Sequencing of the recovered protein resulted in identification of the repressor protein as the sarcosine oxidase α subunit. This was confirmed by expression of the gene encoding the M. loti α subunit of sarcosine oxidase in Escherichia coli. When the expressed peptide was bound to RBD, the gel retardation result was identical to the result obtained with rep2037 from M. loti strain NZP2037.

Spatially isolated reactions in a complex array: using magnetic beads to purify and quantify nucleic acids with digital and quantitative real-time PCR in thousands of parallel microwells

Lab on a Chip ◽  
2020 ◽  
Vol 20 (10) ◽  
pp. 1771-1779 ◽  
Author(s):  
W. Hampton Henley ◽  
Nathan A. Siegfried ◽  
J. Michael Ramsey
Keyword(s):  
Nucleic Acid ◽  
Sample Preparation ◽  
Nucleic Acids ◽  
Real Time ◽  
Real Time Pcr ◽  
Magnetic Beads ◽  
Digital Pcr ◽  
Quantitative Real Time Pcr

Encoded beads carrying primer pairs for nucleic acid targets are used for sample preparation and multiplexed-in-space digital PCR quantification.

scholarly journals Magnetic beads-based nucleic acids extraction in microfluidic chip

2019 ◽  
Vol 1400 ◽  
pp. 033012
Author(s):  
P K Afonicheva ◽  
N A Esikova ◽  
A N Tupik ◽  
A A Evstrapov
Keyword(s):  
Nucleic Acids ◽  
Microfluidic Chip ◽  
Magnetic Beads

scholarly journals Optimization of PCR Purification Using Silica-Coated Magnetic Beads

2020 ◽  
Author(s):  
K.T. Berdimuratova ◽  
A.O Amirgazin ◽  
М.А. Kuibagarov ◽  
V.B. Lutsay ◽  
K.K. Mukanov ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Magnetic Beads ◽  
Molecular Genetic ◽  
Genetic Research ◽  
Buffer System ◽  
Dna Fragments ◽  
Dna Molecules ◽  
Sequencing Technologies ◽  
Pcr Products ◽  
The Cost

Purification of nucleic acids is still an important step in molecular genetic research. The development of whole genome sequencing technologies has increased the requirements for the purity of the nucleic acids used, and also required the selection of DNA fragments by size. Buffer systems that contain PEG/NaCl solutions and silica-coated magnetic beads allow to purify nucleic acids and selectively sorb certain sizes of DNA. In this article, we present a simple protocol for the purification of PCR products with the ability to absorb the required DNA molecules. It was determined that the use of an optimized PEG / NaCl buffer system with magnetic silica gel in a ratio of 1.5: 1 with a PCR product allows to get rid of DNA fragments 100 and less base pairs (bp), as well as other contaminants, while maintaining this is more than 90% of the DNA in solution. The ratio of 0.35: 1 allows for high-affinity sorption of DNA molecules larger than 400 bp. The practical use of the obtained data allows us to improve the quality of sequencing without increasing the cost of research.

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