Long-term room temperature storage of DNA molecules

Biomics ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 552-563
Author(s):  
R.R. Garafutdinov ◽  
A.R. Sakhabutdinova ◽  
A.V. Chemeris

The simplest and most common method of long-term storage of DNA samples at present is the storage of their frozen solutions, which, however, has a number of disadvantages, including the destruction of DNA molecules during freezing and thawing, as well as energy consumption and the likelihood of losing valuable samples in the event of possible accidents. In this regard, long-term storage of DNA samples at room temperature in a dried state is preferable, especially since an even greater increase in the number of stored DNA samples is planned due to the planned preservation of non-biological data in this molecule, which is recognized at the International Economic Forum 2019 among the 10 most important innovative technologies as “DNA Data Storage” of the near future of mankind. Such storage requires the exclusion of hydrolysis and oxidation of DNA molecules under the action of water and reactive oxygen species, which can be achieved by placing DNA in an inert anhydrous atmosphere, including in the presence of additional ingredients in the form of, for example, trehalose, imitating wildlife, since it is known that this simple disaccharide, capable of vitrification, protects a wide range of anhydrobiont organisms from adverse environmental conditions. Currently, there are a number of technologies that provide long-term storage of DNA at room temperature, including those available from commercial sources, but not all problems have yet been solved, which is reflected in this review article.

Author(s):  
A. Sinebryukhova ◽  
A. Shipelova ◽  
E. Darnotuk ◽  
A. Chekanov ◽  
O. Baranova ◽  
...  

The optimal conditions were selected for obtaining homogeneous nanoemulsions (NE) of lipoic acid conjugates (LA-conjugates) based on Pluronic F68 (1,8%) with a particle size not exceeding 400 nm, characterized by 97±2% encapsulation efficiency of substances in nanoparticles (NP). A heterogeneous NE (polydispersity index, PDI>0,3) with the derivative of LA and myo-inositol based on phosphatidylcholine (PC, C = 3 mg/ml) was also obtained consisting of 2 particle fractions: 20–70 nm (27%) and 122–212 nm (73%). The obtained NEs with LA-conjugates based on Pluronic F68 and PC were stable during long-term storage (more than 12 months) at room temperature. The effect of the obtained NEs of LA-conjugates on platelet aggregation (Pt) caused by arachidonic acid (AA) was determined, and a mechanism of their action was proposed.


2020 ◽  
Vol 65 (No. 4) ◽  
pp. 115-123
Author(s):  
Marija Jovičić ◽  
Eva Chmelíková ◽  
Markéta Sedmíková

Sperm cryopreservation is the best technology for long-term storage of the semen. However, the damage of boar spermatozoa by cryopreservation is more severe than in other animal species and a standardized freezing protocol for efficient cryopreservation has not been established yet. Semen quality and freezability vary greatly between breeds as well as between individual boars and even the season. Boar spermatozoa are sensitive to low temperatures; they sustain damage and a high rate of mortality and freezing/thawing the boar semen may strongly impair the sperm function and decrease the semen quality. The freezability of boar semen can be influenced by a cryopreservation procedure, and also by using various additives to freezing and thawing extenders such as antioxidants. In order to obtain acceptable results after thawing the boar semen, it is necessary to combine an optimal amount of additives (glycerol, egg yolk, sugars, antioxidants), cooling and warming velocities.


2019 ◽  
Vol 26 (1) ◽  
Author(s):  
William Ottestad ◽  
Ingrid N. Rognes ◽  
Erlend Skaga ◽  
Cassandra Frisvoll ◽  
Guttorm Haraldsen ◽  
...  

Abstract Background HMGB1 is a mediator of systemic inflammation in sepsis and trauma, and a promising biomarker in many diseases. There is currently no standard operating procedure for pre-analytical handling of HMGB1 samples, despite that pre-analytical conditions account for a substantial part of the overall error rate in laboratory testing. We hypothesized that the considerable variations in reported HMGB1 concentrations and kinetics in trauma patients could be partly explained by differences in pre-analytical conditions and choice of sample material. Methods Trauma patients (n = 21) admitted to a Norwegian Level I trauma center were prospectively included. Blood was drawn in K2EDTA coated tubes and serum tubes. The effects of delayed centrifugation were evaluated in samples stored at room temperature for 15 min, 3, 6, 12, and 24 h respectively. Plasma samples subjected to long-term storage in − 80 °C and to repeated freeze/thaw cycles were compared with previously analyzed samples. HMGB1 concentrations in simultaneously acquired arterial and venous samples were also compared. HMGB1 was assessed by standard ELISA technique, additionally we investigated the suitability of western blot in both serum and plasma samples. Results Arterial HMGB1 concentrations were consistently lower than venous concentrations in simultaneously obtained samples (arterial = 0.60 x venous; 95% CI 0.30–0.90). Concentrations in plasma and serum showed a strong linear correlation, however wide limits of agreement. Storage of blood samples at room temperature prior to centrifugation resulted in an exponential increase in plasma concentrations after ≈6 h. HMGB1 concentrations were fairly stable in centrifuged plasma samples subjected to long-term storage and freeze/thaw cycles. We were not able to detect HMGB1 in either serum or plasma from our trauma patients using western blotting. Conclusions Arterial and venous HMGB1 concentrations cannot be directly compared, and concentration values in plasma and serum must be compared with caution due to wide limits of agreement. Although HMGB1 levels in clinical samples from trauma patients are fairly stable, strict adherence to a pre-analytical protocol is advisable in order to protect sample integrity. Surprisingly, we were unable to detect HMGB1 utilizing standard western blot analysis.


2008 ◽  
Vol 584-586 ◽  
pp. 1039-1044
Author(s):  
Andrey Korotitskiy ◽  
K.E. Inaekyan ◽  
Vladimir Brailovski ◽  
Sergey Prokoshkin

Ti-50.26at.%Ni shape memory alloy samples were subjected to cold rolling (CR) with true strains encompassing from moderate (logarithmic strain e=0.25) to severe (e=2.1) deformation. СR with e = 0.5 and more initiated a partial austenite amorphization. The evaluation of structural changes in the material during its long-term storage was performed using Vickers microhardness (HV) technique. It was shown that during storage at room temperature up to 9 months, microhardness varied following a dome-shaped trend, thus reflecting commonly encountered interaction between two concurrent time-dependent phenomena, the first responsible for the material hardening, and the second, for the material softening. To represent such phenomena, a simple mathematical model was proposed and experimentally validated.


F1000Research ◽  
2017 ◽  
Vol 6 ◽  
pp. 227 ◽  
Author(s):  
Scott Gigante

Oxford Nanopore Technologies' (ONT's) MinION and PromethION long-read sequencing technologies are emerging as genuine alternatives to established Next-Generation Sequencing technologies. A combination of the highly redundant file format and a rapid increase in data generation have created a significant problem both for immediate data storage on MinION-capable laptops, and for long-term storage on lab data servers. We developed Picopore, a software suite offering three methods of compression. Picopore's lossless and deep lossless methods provide a 25% and 44% average reduction in size, respectively, without removing any data from the files. Picopore's raw method provides an 88% average reduction in size, while retaining biologically relevant data for the end-user. All methods have the capacity to run in real-time in parallel to a sequencing run, reducing demand for both immediate and long-term storage space.


Author(s):  
Margarita Ishmuratova ◽  
◽  
Damirzhan Baigarayev ◽  
Saltanat Tleukenova ◽  
Elena Gavrilkova ◽  
...  

This article presents the summarized data on cryopreservation of seeds of the medical plant Nepeta cataria. Cryopreservation is a highly promising method for saving of seed materials, allowing to organize long-term storage without viability loss. The purpose of present work is to optimize conditions of cryopreservation of seed materials of Nepeta cataria. Assessment of seed survival rate in the storage showed a linear decrease in seed viability and energy of germination. After 30 months of storage at the low positive temperature (+5 ºC) in paper pack seed rate decreased to 12.0 % and energy of germination to 11.2 %; after 4 years of storage seeds lost viability. During conduction of research the type of container, condition of thawing, optimal moisture of seeds and cryoprotectants are optimized. The optimal container for cryopreservation in liquid nitrogen was plastic cryo tubes; defrosting at room temperature. The best seed rate is found at moisture 3 %; the best cryoprotectant was glucose, the optimal concentration was 15 %. The result of the research is used for creation of the long-term storage medicinal cultures’ seed bank in the liquid nitrogen.


Vestnik MGTU ◽  
2021 ◽  
Vol 24 (4) ◽  
pp. 441-449
Author(s):  
M. N. Shkolnikova ◽  
V. N. Abbazova

Pumpkin fruits ( Cucurbita spp.) have a number of advantages and high technological potential, thanks to almost universal cultivation in a wide range of agro-climatic conditions, the ability to long-term storage, the content of dietary fibers, pectin and other polysaccharides, carotenoids, polyphenolic substances, vitamins, which cause a wide range of physiological orientation. In the process of researching the composition of local cultivars of pumpkin and the world experience of using Cucurbita spp. in the composition of food products, the need to use this ingredient in beverage recipes has been substantiated. The content of dry substances in the pumpkin samples is from 8.18 % ("Gribovskaya") to 11.6 % ("Orange bush"). The maximum sugar content is distinguished by the varieties "Winter Sweet" (6.87 %) and "Orange Bush" (7.40 %). The content of BAS-antioxidants is (without visible difference depending on the growing region): carotenoids from 1.3 mg/100 g in "Gribovskaya" to 2.0 mg/100 g - "Orange bush"; ascorbic acid - 8.7 mg/100 g in "Rossiyanka" to 14.2 mg/100 g - "Orange bush". Today all parts of the pumpkin fruit are used: the bark is a raw material for the production of feed flour and a substrate for the cultivation of lactobacilli, the seeds are traditionally used to produce pumpkin oil and flour, the pulp of the fruit is a raw material for juice-containing products, purees, carotenoid-containing and polysaccharide extracts, pectin, concentrates of first and second dishes, snack products, pasta, bakery, confectionery and meat products, etc.


2018 ◽  
Vol 29 (1) ◽  
pp. 94-111 ◽  
Author(s):  
Tomás Barranco ◽  
Asta Tvarijonaviciute ◽  
Damián Escribano ◽  
Fernando Tecles ◽  
José J Cerón ◽  
...  

Introduction: In this report, we aimed to examine the stability of various analytes in saliva under different storage conditions. Materials and methods: Alpha-amylase (AMY), cholinesterase (CHE), lipase (Lip), total esterase (TEA), creatine kinase (CK), aspartate aminotransferase (AST), lactate dehydrogenase (LD), lactate (Lact), adenosine deaminase (ADA), Trolox equivalent antioxidant capacity (TEAC), ferric reducing ability (FRAS), cupric reducing antioxidant capacity (CUPRAC), uric acid (UA), catalase (CAT), advanced oxidation protein products (AOPP) and hydrogen peroxide (H2O2) were colorimetrically measured in saliva obtained by passive drool from 12 healthy voluntary donors at baseline and after 3, 6, 24, 72 hours, 7 and 14 days at room temperature (RT) and 4 ºC, and after 14 days, 1, 3 and 6 months at – 20 ºC and – 80 ºC. Results: At RT, changes appeared at 6 hours for TEA and H2O2; 24 hours for Lip, CK, ADA and CUPRAC; and 72 hours for LD, Lact, FRAS, UA and AOPP. At 4 ºC changes were observed after 6 hours for TEA and H2O2; 24 hours for Lip and CUPRAC; 72 hours for CK; and 7 days for LD, FRAS and UA. At – 20 ºC changes appeared after 14 days for AST, Lip, CK and LD; and 3 months for TEA and H2O2. At – 80 ºC observed changes were after 3 months for TEA and H2O2. Conclusions: In short-term storage, the analytes were more stable at 4 ºC than at room temperature, whereas in long-term storage they were more stable at - 80 ºC than at – 20 ºC.


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