scholarly journals Isolation of high-molecular weight (HMW) DNA from Diplonema papillatum v2 (protocols.io.bb5eiq3e)

protocols.io ◽  
2020 ◽  
Author(s):  
Matus Valach
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Hmw Dna

Rapid and efficient method for isolating plant high molecular weight (HMW) DNA with high purity

1999 ◽  
Vol 44 (4) ◽  
pp. 378-380 ◽  
Author(s):  
Fang Qiu ◽  
Jianmin Fu ◽  
Weiwu Xie ◽  
Chuanyou Li ◽  
Bin Wang
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Efficient Method ◽  
High Purity ◽  
Hmw Dna

scholarly journals Isolation of High Molecular Weight DNA from the Model Beetle Tribolium for Nanopore Sequencing

Genes ◽  
2021 ◽  
Vol 12 (8) ◽  
pp. 1114
Author(s):  
Marin Volarić ◽  
Damira Veseljak ◽  
Brankica Mravinac ◽  
Nevenka Meštrović ◽  
Evelin Despot-Slade
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Critical Factor ◽  
Anion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Total Output ◽  
Nanopore Sequencing ◽  
High Molecular Weight Dna ◽  
Long Read ◽  
Hmw Dna

The long-read Nanopore sequencing has been recently applied for assembly of complex genomes and analysis of linear genome organization. The most critical factor for successful long-read sequencing is extraction of high molecular weight (HMW) DNA of sufficient purity and quantity. The challenges associated with input DNA quality are further amplified when working with extremely small insects with hard exoskeletons. Here, we optimized the isolation of HMW DNA from the model beetle Tribolium and tested for use in Nanopore sequencing. We succeeded in overcoming all the difficulties in HMW handling and library preparation that were encountered when using published protocols and commercial kits. Isolation of nuclei and subsequent purification of DNA on an anion-exchange chromatography column resulted in genomic HMW DNA that was efficiently relaxed, of optimal quality and in sufficient quantity for Nanopore MinION sequencing. DNA shearing increased average N50 read values up to 26 kb and allowed us to use a single flow cell in multiple library loads for a total output of more than 13 Gb. Although our focus was on T. castaneum and closely related species, we expect that this protocol, with appropriate modifications, could be extended to other insects, particularly beetles.

Abstract 1704: AC electrokinetic isolation of cell free high molecular weight DNA (CF-HMW DNA) from serum

2012 ◽  
Author(s):  
Jerry Lu ◽  
William Hanna ◽  
Lucas S. Kumosa ◽  
Megha Bhalla ◽  
Veronica W. Cheung ◽  
...  
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
High Molecular Weight Dna ◽  
Hmw Dna

Salting-out Protocol for Extracting HMW Genomic DNA from frozen octocorals v1

2021 ◽  
Author(s):  
Santiago Herrera
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Liquid Nitrogen ◽  
Genome Sequencing ◽  
Genomic Dna ◽  
Starting Material ◽  
Salting Out ◽  
Hmw Dna

Salting-out protocol for extracting high-molecular weight (HMW) genomic DNA from frozen octocorals for genomic methods. Suitable for genome sequencing, RAD-seq and other methods that require HMW DNA. Starting material must be alive when flash-frozen in liquid nitrogen and always stored below -80C.

scholarly journals High molecular weight DNA extraction strategies for long-read sequencing of complex metagenomes

2021 ◽  
Author(s):  
Florian Trigodet ◽  
Karen Lolans ◽  
Emily Fogarty ◽  
Alon Shaiber ◽  
Hilary G. Morrison ◽  
...  
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Dna Extraction ◽  
Cultured Cells ◽  
High Molecular Weight Dna ◽  
Human Tongue ◽  
Long Read ◽  
Hmw Dna ◽  
High Concentrations ◽  
Assembly Performance

AbstractBy offering extremely long contiguous characterization of individual DNA molecules, rapidly emerging long-read sequencing strategies offer comprehensive insights into the organization of genetic information in genomes and metagenomes. However, successful long-read sequencing experiments demand high concentrations of highly purified DNA of high molecular weight (HMW), which limits the utility of established DNA extraction kits designed for short-read sequencing. Challenges associated with input DNA quality intensify further when working with complex environmental samples of low microbial biomass, which requires new protocols that are tailored to study metagenomes with long-read sequencing. Here, we use human tongue scrapings to benchmark six HMW DNA extraction strategies that are based on commercially available kits, phenol-chloroform (PC) extraction, and agarose encasement followed by agarase digestion. A typical end goal of HMW DNA extractions is to obtain the longest possible reads during sequencing, which is often achieved by PC extractions as demonstrated in sequencing of cultured cells. Yet our analyses that consider overall read-size distribution, assembly performance, and the number of circularized elements found in sequencing results suggest that non-PC methods may be more appropriate for long-read sequencing of metagenomes.

Microtubule motors and other maps

1990 ◽  
Vol 48 (3) ◽  
pp. 1-1
Author(s):  
Richard B. Vallee
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Cytoplasmic Dynein ◽  
Organelle Transport ◽  
Structural Components ◽  
Microtubule Associated Proteins ◽  
Microtubule Motors ◽  
Associated Proteins ◽  
The Subject ◽  
Intracellular Motility

Microtubules are involved in a number of forms of intracellular motility, including mitosis and bidirectional organelle transport. Purified microtubules from brain and other sources contain tubulin and a diversity of microtubule associated proteins (MAPs). Some of the high molecular weight MAPs - MAP 1A, 1B, 2A, and 2B - are long, fibrous molecules that serve as structural components of the cytamatrix. Three MAPs have recently been identified that show microtubule activated ATPase activity and produce force in association with microtubules. These proteins - kinesin, cytoplasmic dynein, and dynamin - are referred to as cytoplasmic motors. The latter two will be the subject of this talk.Cytoplasmic dynein was first identified as one of the high molecular weight brain MAPs, MAP 1C. It was determined to be structurally equivalent to ciliary and flagellar dynein, and to produce force toward the minus ends of microtubules, opposite to kinesin.

Studies on the Functionality of Newly Synthesized Fibrinogen after Treatment of Acute Myocardial Infarction with Streptokinase, Increase in the Rate of Fibrinopeptide Release

1993 ◽  
Vol 70 (06) ◽  
pp. 0978-0983 ◽  
Author(s):  
Edelmiro Regano ◽  
Virtudes Vila ◽  
Justo Aznar ◽  
Victoria Lacueva ◽  
Vicenta Martinez ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Acute Myocardial Infarction ◽  
Molecular Weight ◽  
Steady State ◽  
High Molecular Weight ◽  
Coronary Arteries ◽  
Risk Index ◽  
Weight Fraction ◽  
High Molecular Weight Fraction ◽  
Fibrinopeptide A

SummaryIn 15 patients with acute myocardial infarction who received 1,500,000 U of streptokinase, the gradual appearance of newly synthesized fibrinogen and the fibrinopeptide release during the first 35 h after SK treatment were evaluated. At 5 h the fibrinogen circulating in plasma was observed as the high molecular weight fraction (HMW-Fg). The concentration of HMW-Fg increased continuously, and at 20 h reached values higher than those obtained from normal plasma. HMW-Fg represented about 95% of the total fibrinogen during the first 35 h. The degree of phosphorylation of patient fibrinogen increased from 30% before treatment to 65% during the first 5 h, and then slowly declined to 50% at 35 h.The early rates of fibrinopeptide A (FPA) and phosphorylated fibrinopeptide A (FPAp) release are higher in patient fibrinogen than in isolated normal HMW-Fg and normal fibrinogen after thrombin addition. The early rate of fibrinopeptide B (FPB) release is the same for the three fibrinogen groups. However, the late rate of FPB release is higher in patient fibrinogen than in normal HMW-Fg and normal fibrinogen. Therefore, the newly synthesized fibrinogen clots faster than fibrinogen in the normal steady state.In two of the 15 patients who had occluded coronary arteries after SK treatment the HMW-Fg and FPAp levels increased as compared with the 13 patients who had patent coronary arteries.These results provide some support for the idea that an increased synthesis of fibrinogen in circulation may result in a procoagulant tendency. If this is so, the HMW-Fg and FPAp content may serve as a risk index for thrombosis.

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