Navigation Links
Efficient Transfection of Neurospora Crassa

be transfected efficiently without having to weaken the cell wall or create protoplasts.

OBJECTIVE:

To provide a simple, efficient method for transfecting N. crassa conidia with the Eppendorf Electroporator 2510.

EXPERIMENTAL METHOD:

Cell Cultivation and Harvesting
A Neurospora crassa wild type strain was cultured in VM agar with appropriate supplements in a 250 ml Erlenmeyer flask (a 125 ml flask may also be used) [3]. The flask was incubated at 30°C for 3 days, then transferred to room temperature and exposed to fluorescent light for an additional 5 days. It is necessary to use conidia that are at least eight days old.

The conidia were harvested using sterile water. The suspension was transferred to a 50 ml conical tube and centrifuged at 2,500 rpm for 5 minutes to pellet the conidia. After aspirating the supernatant, the pellet was re-suspended in 25 ml of sterile water. The tube was vortexed to bring the conidia pellet into solution and then re-centrifuged at 2,500 rpm for 5 minutes. The conidia pellet was re-suspended in 25 ml of cold 1 M sorbitol (vortexed for re-suspension) and then centrifuged as stated above; this was repeated twice.

The final pellet was re-suspended in 0.5 ml cold sorbitol and transferred to a 2 ml Eppendorf microcentrifuge tube. The concentration of the suspension was determined using a hemacytometer. The optimal concentration is approximately 2.5 x 109/ml. It may be necessary to concentrate or dilute the sample to bring it to optimal concentration.

Electroporation Procedure
The suspension was transferred in 40 µl aliquots to two Eppendorf
'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. Efficiently Insert Unique Restriction Sites into Plasmid Vectors
2. Efficient Cleavage of Fusion Proteins to Yield Native Amino Termini
3. Mammalian Expression Vector for Efficient Cloning of PCR Fragments
4. Efficient Transfection of Neurospora Crassa
5. Efficient Recovery of Ultrapure Plasmid DNA
6. Efficient and Reliable PCR Setup Using Eppendorf MasterMix
7. Efficient DNA transfection of primary CNS neurons using TransMessenger Transfection Reagent
8. Efficient RNAi-mediated gene silencing in neuronal cells using QIAGEN siRNA and TransMessenger Transfection Reagent*
9. Cloning Based on Efficient Three-Fragment Assembly DNA Ligation
10. Efficient and Reliable Linear Amplification of cRNA
11. Efficient Delivery of siRNAs to Human Primary Cells: Electroporation vs. Chemical Transfection
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Efficient Transfection Neurospora Crassa

(Date:7/30/2014)... 30, 2014 With the accelerated ... personal health expenditure as well as the improved ... China diagnostic reagent industry has been developing quickly, ... In 2013, Chinese in vitro diagnostics market size ... from in vitro diagnostic reagents. , Read ...
(Date:7/30/2014)... 2014  Orexigen Therapeutics, Inc. (Nasdaq: OREX ... List of Outstanding Issues (LOI) from the European Medicines ... for the NB32 Marketing Authorization Application (MAA). NB32 (naltrexone ... drug candidate being evaluated for weight loss. ... point were adequately addressed by the Company based in ...
(Date:7/30/2014)... 2014  RXi Pharmaceuticals Corporation (NASDAQ: ... on discovering, developing and commercializing innovative therapies ... technologies, today announced that their third Phase ... reduction of recurrence of hypertrophic scars following ... Logo - http://photos.prnewswire.com/prnh/20130917/NE80755LOGO ...
(Date:7/30/2014)... Immunology Summit-2014 will be ... scientists in the field, which includes prominent personalities ... Western Reserve University School of Medicine, USA, who ... “The effects of recombinant human IL-6 and recombinant ... Biragyn, a Senior investigator at National Institute on ...
Breaking Biology Technology:China Diagnostic Reagent Industry Report, 2013-2016 | Researchmoz 2China Diagnostic Reagent Industry Report, 2013-2016 | Researchmoz 3Orexigen Receives CHMP Day 180 List of Outstanding Issues 2Orexigen Receives CHMP Day 180 List of Outstanding Issues 3RXi Pharmaceuticals Expands Clinical Program with Initiation of Third Phase 2a Study with RXI-109 for Treatment of Hypertrophic Scars 2RXi Pharmaceuticals Expands Clinical Program with Initiation of Third Phase 2a Study with RXI-109 for Treatment of Hypertrophic Scars 3RXi Pharmaceuticals Expands Clinical Program with Initiation of Third Phase 2a Study with RXI-109 for Treatment of Hypertrophic Scars 4RXi Pharmaceuticals Expands Clinical Program with Initiation of Third Phase 2a Study with RXI-109 for Treatment of Hypertrophic Scars 5Unveiling Breakthroughs in Clinical & Cellular Immunology 2Unveiling Breakthroughs in Clinical & Cellular Immunology 3
... way of life at Meriter Hospital , a ... care to southern Wisconsin. Meriter includes a hospital, clinic, ... service. Formed from a merger of Madison General Hospital ... 430 active and associate physicians in over 30 medical ...
... from traditional paper phone bills to SBC eBill ... the "Save a StampHelp Save a Life" campaign, created in ... Inc. will contribute $1 dollar for every SBC customer that ... 15, 2004. , ,In 2004, an estimated 216,000 new cases ...
... Despite a two-year-plus economic recovery in the United States, ... which source you cite) have mysteriously failed to appear ... they not been created as expected? The list of ... recovery, uncertainty about the future, terrorism and the war ...
Cached Biology Technology:Meriter IT team builds project performance through internal relationships 2Meriter IT team builds project performance through internal relationships 3Meriter IT team builds project performance through internal relationships 4Gone missing: Today jobs, tomorrow workers? 2Gone missing: Today jobs, tomorrow workers? 3
(Date:7/30/2014)... (125I) in cancer treatment has been shown to ... critical for neural transmission between the peripheral and ... from Institute of Radiation Medicine, Chinese Academy of ... implanted into rat dorsal root ganglia (DRG) to ... different radioactivity (0, 14.8, 29.6 MBq) were implanted ...
(Date:7/30/2014)... into a multitude of cells types. Researchers would like to ... of mature cells that make up the organs and other ... be long chains of sugars that dangle from proteins on ... of California, San Diego, has created synthetic molecules that can ... easily manipulated to direct the process, they report in the ...
(Date:7/30/2014)... SAN JOSE, Calif. , July 30, 2014 ... industry-academia partnership with the United States Air Force ... project proposed by Binghamton ... for human performance monitoring (HPM). The $425,000 project, ... and electronics packaging firm i3 Electronics, Inc. ( ...
Breaking Biology News(10 mins):Nano-Bio Manufacturing Consortium Awards Funding to Binghamton University for Development of Electronics and Biometric Sensor Platforms 2Nano-Bio Manufacturing Consortium Awards Funding to Binghamton University for Development of Electronics and Biometric Sensor Platforms 3
... life," but does today,s unprecedented level of dietary variety ... it might. According to ASN Spokesperson Shelley ... eating, itself- can trigger release of various brain chemicals, ... with drug addiction and withdrawal. And, as can happen ...
... Populations of spur-thighed tortoises ( Testudo graeca ), a species ... fires if outbreaks occur once every three decades or more. ... each fire. These are the results of a study by ... fire in the Sierra de la Carrasquilla mountains in Murcia ...
... is available in German . They live their lives ... upright position, sloths spend most of their lives hanging in trees ... slowly. Very slowly. But why are sloths so ,lazy,? And how ... lifestyle in the course of evolution? Zoologists of the Friedrich Schiller ...
Cached Biology News:Does food act physiologically like a 'drug of choice' for some? 2Tortoise populations can withstand fires every 30 years 2The decoding of slowness 2
... The Sciclone ALH 4000 workstation is ... groups seeking to introduce automated liquid handling ... labs that need to support routine applications ... automated and,integrated for walk-away automation even ...
... ECACC is able to accept cell lines produced ... The cell lines must be accompanied by a ... accordance with the requirements of cGMP and that ... bacteria or fungi. What are the benefits of ...
... Affinity Life Sciences provides cost effective cell ... proteins or monoclonal antibodies. Quality at a ... whether you require research or commercial product, ... production methods such as roller bottles, hollow ...
... The Human Genetic Cell Bank Service ... ECACC is based on Epstein-Barr Virus (EBV) ... resulting in the production of lymphoblastoid cell ... banked, providing a renewable and expandable source ...
Biology Products: