scholarly journals A versatile bulk electrotransfection protocol for mouse embryonic fibroblast and iPS cells

2019 ◽  
Author(s):  
Shahin Eghbalsaied ◽  
Iqbal Hyder ◽  
Wilfried A. Kues

AbstractA square-wave pulsing protocol was developed using OptiMEM-GlutaMAX for high efficient transfection of mouse embryonic fibroblast (MEF) and induced pluripotency stem (iPS) cells. An electrotransfection efficiency of > 95% was repeated for both MEF and iPS cells using reporter-encoding plasmids. The protocol was very efficient for plasmid size ranging from 6.2 to 13.5 kb. A high rate of targeted gene knockout (> 95 %) was produced in Venus transgenic cells using indels formation. Targeted deletions in the Venus transgene were performed by co-electroporation of two gRNA-encoding plasmids. In conclusion, this plasmid electrotransfection protocol is straight-forward, cost-effective, and efficient for CRISPRing mouse primary cells.

ChemBioChem ◽  
2012 ◽  
Vol 14 (1) ◽  
pp. 73-82 ◽  
Author(s):  
Noriko Nagahori ◽  
Tadashi Yamashita ◽  
Maho Amano ◽  
Shin-Ichiro Nishimura

Author(s):  
Y. Anggoro

The Belida field is an offshore field located in Block B of Indonesia’s South Natuna Sea. This field was discovered in 1989. Both oil and gas bearing reservoirs are present in the Belida field in the Miocene Arang, Udang and Intra Barat Formations. Within the middle Arang Formation, there are three gas pay zones informally referred to as Beta, Gamma and Delta. These sand zones are thin pay zones which need to be carefully planned and economically exploited. Due to the nature of the reservoir, sand production is a challenge and requires downhole sand control. A key challenge for sand control equipment in this application is erosion resistance without inhibiting productivity as high gas rates and associated high flow velocity is expected from the zones, which is known to have caused sand control failure. To help achieve a cost-effective and easily planned deployment solution to produce hydrocarbons, a rigless deployment is the preferred method to deploy downhole sand control. PSD analysis from the reservoir zone suggested from ‘Industry Rules of Thumb’ a conventional gravel pack deployment as a means of downhole sand control. However, based on review of newer globally proven sand control technologies since adoption of these ‘Industry Rules of Thumb’, a cost-effective solution could be considered and implemented utilizing Ceramic Sand Screen technology. This paper will discuss the successful application at Block B, Natuna Sea using Ceramic Sand Screens as a rigless intervention solution addressing the erosion / hot spotting challenges in these high rate production zones. The erosion resistance of the Ceramic Sand Screen design allows a deployment methodology directly adjacent to the perforated interval to resist against premature loss of sand control. The robust ceramic screen design gave the flexibility required to develop a cost-effective lower completion deployment methodology both from a challenging make up in the well due to a restrictive lubricator length to the tractor conveyancing in the well to land out at the desired set depth covering the producing zone. The paper will overview the success of multi-service and product supply co-operation adopting technology enablers to challenge ‘Industry Rules of Thumb’ replaced by rigless reasoning as a standard well intervention downhole sand control solution where Medco E&P Natuna Ltd. (Medco E&P) faces sand control challenges in their high deviation, sidetracked well stock. The paper draws final attention to the hydrocarbon performance gain resulting due to the ability for choke free production to allow drawing down the well at higher rates than initially expected from this zone.


2013 ◽  
Vol 8 (3-4) ◽  
pp. 469-478 ◽  
Author(s):  
Sandip S. Magdum ◽  
Gauri P. Minde ◽  
Upendra S. Adhyapak ◽  
V. Kalyanraman

The aim of this work was to optimize the biodegradation of polyvinyl alcohol (PVA) containing actual textile wastewater for a sustainable treatment solution. The isolated microbial consortia of effective PVA degrader namely Candida Sp. and Pseudomonas Sp., which were responsible for symbiotic degradation of chemical oxidation demand (COD) and PVA from desizing wastewater. In the process optimization, the maximum aeration was essential to achieve a high degradation rate, where as stirring enhances further degradation and foam control. Batch experiments concluded with the need of 16 lpm/l and 150 rpm of air and stirring speed respectively for high rate of COD and PVA degradation. Optimized process leads to 2 days of hydraulic retention time (HRT) with 85–90% PVA degradation. Continuous study also confirmed above treatment process optimization with 85.02% of COD and 90.3% of PVA degradation of effluent with 2 days HRT. This study gives environment friendly and cost effective solution for PVA containing textile wastewater treatment.


2001 ◽  
Vol 44 (8) ◽  
pp. 15-25 ◽  
Author(s):  
J.B. van Lier ◽  
F.P. van der Zee ◽  
N.C.G. Tan ◽  
S. Rebac ◽  
R. Kleerebezem

Anaerobic wastewater treatment (AnWT) is considered as the most cost-effective solution for organically polluted industrial waste streams. Particularly the development of high-rate systems, in which hydraulic retention times are uncoupled from solids retention times, has led to a world-wide acceptance of AnWT. In the last decade up to the present, the application potentials of AnWT are further explored. Research shows the feasibility of anaerobic reactors under extreme conditions, such as low and high temperatures. Also toxic and/or recalcitrant wastewaters, that were previously believed not to be suitable for anaerobic processes, are now effectively treated. The recent advances are made possible by adapting the conventional anaerobic high-rate concept to the more extreme conditions. Staged anaerobic reactor concepts show advantages under non-optimal temperature conditions as well as during the treatment of chemical wastewater. In other situations, a staged anaerobic - aerobic approach is required for biodegradation of specific pollutants, e.g. the removal of dyes from textile processing wastewaters. The current paper illustrates the benefits of reactor staging and the yet un-exploited potentials of high-rate AnWT.


2002 ◽  
Vol 45 (10) ◽  
pp. 321-328 ◽  
Author(s):  
H.J. Gijzen

After the discovery of methane gas by Alessandro Volta in 1776, it took about 100 years before anaerobic processes for the treatment of wastewater and sludges were introduced. The development of high rate anaerobic digesters for the treatment of sewage and industrial wastewater took until the nineteen-seventies and for solid waste even till the nineteen-eighties. All digesters have in common that they apply natural anaerobic consortia of microorganisms for degradation and transformation processes. In view of this, it could be rewarding to evaluate the efficiency of natural ecosystems for their possible application. Examples of high rate anaerobic natural systems include the forestomach of ruminants and the hindgut of certain insects, such as termites and cockroaches. These “natural reactors” exhibit volumetric methane production rates as high as 35 l/l.d. The development of anaerobic reactors based on such natural anaerobic systems could produce eco-technologies for the effective management of a wide variety of solid wastes and industrial wastewater. Important limitations of anaerobic treatment of domestic sewage relate to the absence of nutrient and pathogen removal. A combination of anaerobic pre-treatment followed by photosynthetic post-treatment is proposed for the effective recovery of energy and nutrients from sewage. This eco-technology approach is based on the recognition that the main nutrient assimilating capacity is housed in photosynthetic plants. The proposed anaerobic-photosynthetic process is energy efficient, cost effective and applicable under a wide variety of rural and urban conditions. In conclusion: a natural systems approach towards waste management could generate affordable eco-technologies for effective treatment and resource recovery.


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