scholarly journals Comparison of Swabbing Solution Volume and gDNA Extraction Kits on DNA Recovery from Rigid Surface

2020 ◽  
Vol 60 (2) ◽  
pp. 206-213
Author(s):  
Yeon Jeong Ok ◽  
Hee Sang You ◽  
Song Hee Lee ◽  
Ho Joong Sung ◽  
Hee-Gyoo Kang ◽  
...  
2019 ◽  
Vol 161 ◽  
pp. 12-17 ◽  
Author(s):  
Hee Sang You ◽  
Song Hee Lee ◽  
Yeon Jeong Ok ◽  
Hee-Gyu Kang ◽  
Ho Jung Sung ◽  
...  

mSphere ◽  
2019 ◽  
Vol 4 (5) ◽  
Author(s):  
Katja Engel ◽  
Sara Coyotzi ◽  
Melody A. Vachon ◽  
Jennifer R. McKelvie ◽  
Josh D. Neufeld

ABSTRACT Bentonite clay is an integral component of the engineered barrier system of deep geological repositories (DGRs) that are planned for the long-term storage of high-level radioactive waste. Although nucleic acid extraction and analysis can provide powerful qualitative and quantitative data reflecting the presence, abundance, and functional potential of microorganisms within DGR materials, extraction of microbial DNA from bentonite clay is challenging due to the low biomass and adsorption of nucleic acids to the charged clay matrix. In this study, we used quantitative PCR, gel fingerprinting, and high-throughput sequencing of 16S rRNA gene amplicons to assess DNA extraction efficiency from natural MX-80 bentonite and the same material “spiked” with Escherichia coli genomic DNA. Extraction protocols were tested without additives and with casein and phosphate as blocking agents. Although we demonstrate improved DNA recovery by blocking agents at relatively high DNA spiking concentrations, at relatively low spiking concentrations, we detected a high proportion of contaminant nucleic acids from blocking agents that masked sample-specific microbial profile data. Because bacterial genomic DNA associated with casein preparations was insufficiently removed by UV treatment, casein is not recommended as an additive for DNA extractions from low-biomass samples. Instead, we recommend a kit-based extraction protocol for bentonite clay without additional blocking agents, as tested here and validated with multiple MX-80 bentonite samples, ensuring relatively high DNA recoveries with minimal contamination. IMPORTANCE Extraction of microbial DNA from MX-80 bentonite is challenging due to low biomass and adsorption of nucleic acid molecules to the charged clay matrix. Blocking agents improve DNA recovery, but their impact on microbial community profiles from low-biomass samples has not been characterized well. In this study, we evaluated the effect of casein and phosphate as blocking agents for quantitative recovery of nucleic acids from MX-80 bentonite. Our data justify a simplified framework for analyzing microbial community DNA associated with swelling MX-80 bentonite samples within the context of a deep geological repository for used nuclear fuel. This study is among the first to demonstrate successful extraction of DNA from Wyoming MX-80 bentonite.


2017 ◽  
Vol 69 (6) ◽  
pp. 1443-1450 ◽  
Author(s):  
M.P. Campos ◽  
M.F. Madeira ◽  
D.A. Silva ◽  
M.S. Solcà ◽  
O.M. Espíndola ◽  
...  

ABSTRACT The purpose of the present work was to evaluate the accuracy of quantitative polymerase chain reaction (qPCR) performed on samples of fresh frozen tissue (FT) and formalin-fixed, paraffin-embedded (FFPE) healthy skin. This is a validation study conducted with samples from 46 dogs from an endemic area in Brazil. After sample collection, DNA extractions were conducted using commercial kits and qPCR was oriented to kinetoplast DNA (kDNA) targets of the Leishmania infantum species. The results obtained for the FFPE samples showed 63.6% sensitivity and 77.1% specificity, whereas those obtained for the FT samples showed 100% and 48.6%, respectively. Poor agreement was observed for the results of the qPCR technique with FT and FFPE samples. Our results suggest freezing as the most suitable conservation method for the formation of sample databases considering DNA recovery


1985 ◽  
Vol 148 (2) ◽  
pp. 527-532 ◽  
Author(s):  
Bjørn Torger Stokke ◽  
Arne Mikkelsen ◽  
Arnljot Algsaeter

Author(s):  
Patrick McLaughlin ◽  
Christian Hopkins ◽  
Eliot Springer ◽  
Mechthild Prinz

Polymers ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 311 ◽  
Author(s):  
Hammad R. Khalid ◽  
Iqra Choudhry ◽  
Daeik Jang ◽  
Nadir Abbas ◽  
M. Salman Haider ◽  
...  

Flexible electronic devices have gained significant interest due to their different potential applications. Herein, we report highly flexible, stretchable, and sensitive sensors made of sprayed CNT layer, sandwiched between two polymer layers. A facile fabrication process was employed in which the CNT solution was directly sprayed onto a patterned bottom polymer layer, above which a second polymer layer was casted to get a sandwiched composite structure. Varying amounts of CNT solution (i.e., 10, 25, 40, 70, and 100 mL) were sprayed to get conductive CNT layers of different thicknesses/densities. The physical characteristics of the conductive CNT layers were studied through SEM and optical images. The starting electrical resistance values (without strain) as well as the changes in electrical resistance against human body motions were monitored. The synthesized samples exhibited good response against finger and wrist bending. The conductivity of the samples increased with increase of CNT solution volume while the sensitivity followed the inverse relation, suggesting that the sensors with controlled sensitivity could be fabricated for targeted strain ranges using the proposed method.


1984 ◽  
Vol 106 (1) ◽  
pp. 26-34 ◽  
Author(s):  
Bharat Bhushan

The statistical analysis of the real area of contact proposed by Greenwood and Williamson is revisited. General and simplified equations for the mean asperity real area of contact, number of contacts, total real area of contact, and mean real pressure as a function of apparent pressure for the case of elastic junctions are presented. The critical value of the mean asperity pressure at which plastic flow starts when a polymer contacts a hard material is derived. Based on this, conditions of elastic and plastic junctions for polymers are defined by a “polymer” plasticity index, Ψp which depends on the complex modulus, Poisson’s ratio, yield strength, and surface topography. Calculations show that most dynamic contacts that occur in a computer-magnetic tape are elastic, and the predictions are supported by experimental evidence. Tape wear in computer applications is small and decreases Ψp by less than 10 percent. The theory presented here can also be applied to rigid and floppy disks.


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