High-throughput automation design considerations for biotechnology processes involving RNA purification protocols using multi-centrifuge bioseparation steps

2012 ◽  
Vol 28 (3) ◽  
pp. 285-293 ◽  
Author(s):  
Aura-Maria Cardona ◽  
Zvi Roth ◽  
Chingping Han
Keyword(s):  
High Throughput ◽  
Design Considerations ◽  
Rna Purification

scholarly journals A simple method for in situ, multiplexed measurement of RNA degradation by flow cytometry

2021 ◽  
Author(s):  
Jaya Rammohan ◽  
Nina Alperovich ◽  
Bin Shao ◽  
David Ross
Keyword(s):  
Flow Cytometry ◽  
High Throughput ◽  
Biological Function ◽  
Rna Degradation ◽  
Rna Sequences ◽  
Simple Method ◽  
Rna Purification ◽  
Degradation Rates ◽  
Cellular Context

RNA degradation plays a major role in cellular function, but current methods for measuring RNA degradation require RNA purification or are low throughput. Here we show how a flow-FISH assay can be used for high-throughput, in situ measurement of RNA degradation without RNA purification. We demonstrate how this approach can be used to simultaneously measure RNA degradation rates of different RNA sequences in a single assay and explore how the assay can be used to examine the effect of cellular context on RNA degradation rates. This assay will be generally useful to quantitatively measure how natural and engineered biological function depends on RNA half-life.

scholarly journals Simultaneous RNA purification and size selection using on-chip isotachophoresis with an ionic spacer

Lab on a Chip ◽  
2019 ◽  
Vol 19 (16) ◽  
pp. 2741-2749 ◽  
Author(s):  
Crystal M. Han ◽  
David Catoe ◽  
Sarah A. Munro ◽  
Ruba Khnouf ◽  
Michael P. Snyder ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Sequencing ◽  
Rna Extraction ◽  
Size Selection ◽  
Rna Purification ◽  
On Chip

We present an on-chip method that achieves simultaneous RNA extraction and size selection, and demonstrate its compatibility with high-throughput sequencing.

scholarly journals A simple, safe and sensitive method for SARS-CoV-2 inactivation and RNA extraction for RT-qPCR

2020 ◽  
Author(s):  
Lelde Kalnina ◽  
Àngels Mateu-Regué ◽  
Stephanie Oerum ◽  
Annemette Hald ◽  
Jan Gerstoft ◽  
...  
Keyword(s):  
High Throughput ◽  
Diagnostic Tests ◽  
Rna Extraction ◽  
High Sensitivity ◽  
Viral Rna ◽  
Extraction Protocol ◽  
Rna Purification ◽  
High Throughput Detection ◽  
Very High ◽  
Sensitive Method

ABSTRACTThe SARS-CoV-2 pandemic has created an urgent need for large amounts of diagnostic tests to detect viral RNA, which commercial suppliers are increasingly unable to deliver. In addition to the lack of availability, the current methods do not always fully inactivate the virus. Together, this calls for the development of safer methods for extraction and detection of viral RNA from patient samples that utilise readily available reagents and equipment present in most standard laboratories. We present a rapid and straightforward RNA extraction protocol for inactivating the SARS-CoV-2 virus that uses standard lab reagents. This protocol expands analysis capacity as the inactivated samples can be used in RT-qPCR detection tests at laboratories not otherwise classified for viral work. The method circumvents the need for commercial RNA purification kits, takes about 30 minutes from swab to PCR-ready viral RNA, and enables downstream detection of SARS-CoV-2 by RT-qPCR with very high sensitivity (~4 viral RNA copies per RT-qPCR). In summary, we present a rapid, safe and sensitive method for high-throughput detection of SARS-CoV-2, that can be conducted in any laboratory equipped with a qPCR machine.

scholarly journals Versatile and flexible microfluidic qPCR test for high-throughput SARS-CoV-2 and cellular response detection in nasopharyngeal swab samples

2020 ◽  
Author(s):  
Julien Fassy ◽  
Caroline Lacoux ◽  
Sylvie Leroy ◽  
Latifa Noussair ◽  
Sylvain Hubac ◽  
...  
Keyword(s):  
High Throughput ◽  
Cellular Response ◽  
Rna Extraction ◽  
Reference Method ◽  
Proteinase K ◽  
Clinical Samples ◽  
Nasopharyngeal Swab ◽  
Rna Purification ◽  
Large Populations ◽  
Reaction Volumes

AbstractThe emergence and quick spread of SARS-CoV-2 has pointed at a low capacity response for testing large populations in many countries, in line of material, technical and staff limitations. The traditional RT-qPCR diagnostic test remains the reference method and is by far the most widely used test. These assays are limited to a couple of probe sets, require large sample PCR reaction volumes, along with an expensive and time-consuming RNA extraction steps. Here we describe a quantitative nanofluidic assay that overcomes some of these shortcomings, based on the Biomark instrument from Fluidigm. This system offers the possibility of performing 4608 qPCR end-points in a single run, equivalent to 192 clinical samples combined with 12 pairs of primers/probe sets in duplicate, thus allowing the monitoring in addition to SARS-CoV-2 probes of other pathogens and/or host cellular responses (virus receptors, response markers, microRNAs). Its 10 nL range volume is compatible with sensitive and reproducible reactions that can be easily and cost-effectively adapted to various RT-qPCR configurations and sets of primers/probe. Finally, we also evaluated the use of inactivating lysis buffers composed of various detergents in the presence or absence of proteinase K to assess the compatibility of these buffers with a direct reverse transcription enzymatic step and we propose several procedures, bypassing the need for RNA purification. We advocate that the combined utilization of an optimized processing buffer and a high-throughput real-time PCR device would contribute to improve the turn-around-time to deliver the test results to patients and increase the SARS-CoV-2 testing capacities.

Morphological behavior of compatibilized ternary blends prepared by mechanical mixing

1993 ◽  
Vol 51 ◽  
pp. 1200-1201
Author(s):  
S.D. Smith ◽  
R.J. Spontak ◽  
D.H. Melik ◽  
S.M. Buehler ◽  
K.M. Kerr ◽  
...  
Keyword(s):  
Interfacial Adhesion ◽  
Diblock Copolymers ◽  
Ternary Blends ◽  
Mechanical Mixing ◽  
Design Considerations ◽  
Covalently Bonded ◽  
Homopolymer Blends ◽  
Emulsifying Agent ◽  
Surface Active ◽  
Low Entropy

When blended together, homopolymers A and B will normally macrophase-separate into relatively large (≫1 μm) A-rich and B-rich phases, between which exists poor interfacial adhesion, due to a low entropy of mixing. The size scale of phase separation in such a blend can be reduced, and the extent of interfacial A-B contact and entanglement enhanced, via addition of an emulsifying agent such as an AB diblock copolymer. Diblock copolymers consist of a long sequence of A monomers covalently bonded to a long sequence of B monomers. These materials are surface-active and decrease interfacial tension between immiscible phases much in the same way as do small-molecule surfactants. Previous studies have clearly demonstrated the utility of block copolymers in compatibilizing homopolymer blends and enhancing blend properties such as fracture toughness. It is now recognized that optimization of emulsified ternary blends relies upon design considerations such as sufficient block penetration into a macrophase (to avoid block slip) and prevention of a copolymer multilayer at the A-B interface (to avoid intralayer failure).

Theoretical And Practical Design Considerations for Ultra-High Resolution Electron Lenses

1980 ◽  
Vol 38 ◽  
pp. 266-269
Author(s):  
Y. Harada ◽  
K. Tsuno ◽  
Y. Arai
Keyword(s):  
Optical Properties ◽  
High Resolution ◽  
Chromatic Aberration ◽  
Point Of View ◽  
Pole Piece ◽  
Axial Field ◽  
Design Considerations ◽  
Resolution Electron ◽  
Practical Design ◽  
Peak Flux

Magnetic objective lenses, from the point of view of pole piece geometry, can he roughly classified into two types, viz., symmetrical and asymmetrical. In the case of the former, the optical properties have been calculated by several authors1-3) and the results would appear to suggest that, in order to reduce the spherical and chromatic aberration coefficients, Cs and Cc, it is necessary to decrease the half-width value of the axial field distribution and to increase the peak flux density. The expressions for either minimum Cs or minimum Cc were presented in the form of ‘universal’ curves by Mulvey and Wallington4).

Sign in / Sign up

Export Citation Format

Share Document