Stop Buffer for Cell Harvesting

2017 ◽  
Vol 2017 (2) ◽  
pp. pdb.rec092932
Keyword(s):  
2017 ◽  
Vol 83 (9) ◽  
Author(s):  
Adam Jordan ◽  
Jenna Chandler ◽  
Joshua S. MacCready ◽  
Jingcheng Huang ◽  
Katherine W. Osteryoung ◽  
...  

ABSTRACT Cyanobacteria are emerging as alternative crop species for the production of fuels, chemicals, and biomass. Yet, the success of these microbes depends on the development of cost-effective technologies that permit scaled cultivation and cell harvesting. Here, we investigate the feasibility of engineering cell morphology to improve biomass recovery and decrease energetic costs associated with lysing cyanobacterial cells. Specifically, we modify the levels of Min system proteins in Synechococcus elongatus PCC 7942. The Min system has established functions in controlling cell division by regulating the assembly of FtsZ, a tubulin-like protein required for defining the bacterial division plane. We show that altering the expression of two FtsZ-regulatory proteins, MinC and Cdv3, enables control over cell morphology by disrupting FtsZ localization and cell division without preventing continued cell growth. By varying the expression of these proteins, we can tune the lengths of cyanobacterial cells across a broad dynamic range, anywhere from an ∼20% increased length (relative to the wild type) to near-millimeter lengths. Highly elongated cells exhibit increased rates of sedimentation under low centrifugal forces or by gravity-assisted settling. Furthermore, hyperelongated cells are also more susceptible to lysis through the application of mild physical stress. Collectively, these results demonstrate a novel approach toward decreasing harvesting and processing costs associated with mass cyanobacterial cultivation by altering morphology at the cellular level. IMPORTANCE We show that the cell length of a model cyanobacterial species can be programmed by rationally manipulating the expression of protein factors that suppress cell division. In some instances, we can increase the size of these cells to near-millimeter lengths with this approach. The resulting elongated cells have favorable properties with regard to cell harvesting and lysis. Furthermore, cells treated in this manner continue to grow rapidly at time scales similar to those of uninduced controls. To our knowledge, this is the first reported example of engineering the cell morphology of cyanobacteria or algae to make them more compatible with downstream processing steps that present economic barriers to their use as alternative crop species. Therefore, our results are a promising proof-of-principle for the use of morphology engineering to increase the cost-effectiveness of the mass cultivation of cyanobacteria for various sustainability initiatives.


Blood ◽  
2017 ◽  
Vol 130 (14) ◽  
pp. 1679-1682 ◽  
Author(s):  
Hira Mian ◽  
Theodore E. Warkentin ◽  
Jo-Ann I. Sheppard ◽  
Anna MacDonald ◽  
Lori-Ann Linkins ◽  
...  
Keyword(s):  

2001 ◽  
Vol 10 (1) ◽  
pp. 17-18
Author(s):  
José R. Borbolla ◽  
Alejandro Nájera ◽  
Manuel A. López-Hernández ◽  
Mauricio González ◽  
Alberto Siller ◽  
...  

2022 ◽  
Author(s):  
Thibault Voeltzel ◽  
Gaëlle Fossard ◽  
Michaël Degaud ◽  
Kevin Geistlich ◽  
Nicolas Gadot ◽  
...  

We provide an easy to access microphysiological standardized system approaching the human bone marrow complexity to a first level of analysis by in situ imaging or by viable cell harvesting of processes taking place within this ecosystem.


2014 ◽  
Vol 778-780 ◽  
pp. 1030-1033 ◽  
Author(s):  
Sei Hyung Ryu ◽  
Craig Capell ◽  
Charlotte Jonas ◽  
Michael J. O'Loughlin ◽  
Jack Clayton ◽  
...  

A 1 cm x 1 cm 4H-SiC N-IGBT exhibited a blocking voltage of 20.7 kV with a leakage current of 140 μA, which represents the highest blocking voltage reported from a semiconductor power switching device to this date. The device used a 160 μm thick drift layer and a 1 μm thick Field-Stop buffer layer, and showed a VF of 6.4 V at an IC of 20 A, and a differential Ron,sp of 28 mΩ-cm2. Switching measurements with a supply voltage of 8 kV were performed, and a turn-off time of 1.1 μs and turn-off losses of 10.9 mJ were measured at 25°C, for a 8.4 mm x 8.4 mm device with 140 μm drift layer and 2 μm F-S buffer layer. The turn-off losses were reduced by approximately 50% by using a 5 μm F-S buffer layer. A 55 kW, 1.7 kV to 7 kV boost converter operating at 5 kHz was demonstrated using the 4H-SiC N-IGBT, and an efficiency value of 97.8% was reported.


2022 ◽  
Author(s):  
Jason Nguyen ◽  
Rebecca Hickman ◽  
Tracy Lee ◽  
Natalie Prystajecky ◽  
John Tyson

This procedure provides instructions on how to prepare DNA libraries for whole genome sequencing on an Illumina MiSeq or NextSeq using Illumina’s DNA Prep Library Preparation Kit scaled to half reaction volumes with modifications to the post-PCR procedures; tagmentation stop buffer and associated washes are removed and libraries are pooled post PCR then a single size selection is performed. This protocol is used to sequence SARS-CoV-2 using the cDNA/PCR protocol: https://dx.doi.org/10.17504/protocols.io.b3viqn4e


Blood ◽  
2000 ◽  
Vol 95 (9) ◽  
pp. 2993-2994 ◽  
Author(s):  
Jan Storek ◽  
Monja A. Dawson ◽  
David G. Maloney

2020 ◽  
Vol 6 (4) ◽  
pp. 2020-2028 ◽  
Author(s):  
Xiaojun Long ◽  
Yang Yi ◽  
Xiaozhao Wang ◽  
Xiyue Duan ◽  
Yuan Sun ◽  
...  

2019 ◽  
Vol 36 (7) ◽  
pp. 1715-1729 ◽  
Author(s):  
Stacey Kowal ◽  
Eliza Kruger ◽  
Pinar Bilir ◽  
James H. Holmes ◽  
William Hickerson ◽  
...  

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