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:1/15/2014)... MINNEAPOLIS , Jan. 15, 2014 ­ RedBrick ... and behavior change technology, today announces that EmblemHealth ... insurer and wellness company, is now providing the RedBrick ... digital coaching program, to all of its members. EmblemHealth ...
(Date:1/15/2014)... 2014 A study has been launched to ... 1 track could help to tackle the problem of obesity. ... (a GP surgery based in Stowmarket) and academics at University ... Telemetry technology, which is inspired by equipment used to ...
(Date:1/15/2014)... (PRWEB) January 15, 2014 Two champions ... continued their sponsorship of an annual competition for middle ... and access to, innovative STEM study. The competition presents ... TEAMS: Tests of Engineering Aptitude, Mathematics, and Sciences ...
(Date:1/15/2014)... DTS Language Services, Inc . is ... for Life Science organizations who need document translations. Clients ... of their documents in advance with a selection of nearly ... translations, often a critical factor in clinical and scientific fields, ...
Breaking Biology Technology:EmblemHealth Selects RedBrick Health to Power its Next-Generation Health Engagement and Behavior Change System 2EmblemHealth Selects RedBrick Health to Power its Next-Generation Health Engagement and Behavior Change System 3Formula 1 Technology Tackles Obesity in Unique Healthcare Partnership 2Formula 1 Technology Tackles Obesity in Unique Healthcare Partnership 3Formula 1 Technology Tackles Obesity in Unique Healthcare Partnership 4Technology Student Association Partners with Leaders in STEM Education Advocacy for Annual Competition 2Technology Student Association Partners with Leaders in STEM Education Advocacy for Annual Competition 3Technology Student Association Partners with Leaders in STEM Education Advocacy for Annual Competition 4Technology Student Association Partners with Leaders in STEM Education Advocacy for Annual Competition 5DTS Improves Efficiency for Life Science Document Translations 2
... AlphaBio Centrix, a Nevada-based biotech corporation, announced this week its ... is a term used in programming wearable TheraEX (TM) ... energy field that surrounds all life forms. ... fact, in its basic form, it embodies seven major layers ...
... VILLAHERMOSA, Mexico, March 29, 2012  An advanced mobile bio-containment ... Tabasco to provide bio-surveillance of disease outbreaks. The US-built ... and is the first unit of its kind in ... This lab represents a new generation of technology ...
... Texas, March 28, 2012   Luminex Corporation ... Dr. Amy Altman, vice president of Biodefense will ... Conference , taking place March 27-29, 2012 in Denver, Colorado. ... of Defense,s Joint Program Executive Office for ...
Cached Biology Technology:AlphaBio Centrix, LLC Announces a New Discovery for Programming Energetic Patches 2US Lab Deployed to Boost Mexico's Response to Outbreaks of Infectious Disease 2Luminex Corporation to Speak at Joint Department of Defense and Environmental Protection Agency 5th National Bio-Threat Conference 2Luminex Corporation to Speak at Joint Department of Defense and Environmental Protection Agency 5th National Bio-Threat Conference 3Luminex Corporation to Speak at Joint Department of Defense and Environmental Protection Agency 5th National Bio-Threat Conference 4
(Date:4/17/2014)... hit on a novel method to help kidney ... most effective treatment possible., Kidney stones represent a ... world. If left untreated, apart from being particularly ... other complications. In many patients treated successfully, stone ... effective pathological approach to diagnosis and treatment needs ...
(Date:4/17/2014)... a stark warning on the possible effects of gases ... 32 times that of carbon dioxide. Now a team ... as fully regenerable electron acceptors which helps explain why ... of being released to the atmosphere. However, there are ... enter into a vicious cycle to release large amounts ...
(Date:4/17/2014)... detected new early-warning signs of the potential loss of ... implications for the diagnosis and treatment of diabetic retinopathy, ... , "We had not expected to see such striking ... Ann Elsner, professor and associate dean in the IU ... "We set out to study the early signs, in ...
Breaking Biology News(10 mins):Methane climate change risk suggested by proof of redox cycling of humic substances 2New technique detects microscopic diabetes-related eye damage 2New technique detects microscopic diabetes-related eye damage 3
... summer temperatures could mean some parts of southern England are ... according to a new book launched today (26 May 2008). ... College London, claims that if average summer temperatures in the ... of Hampshire and the Severn valley, which currently contain many ...
... mulberry leaves without feeling ill, and researchers have come ... special digestive enzyme that is not affected by mulberrys ... amount of alkaloids that inhibit enzymes that break down ... one type of sucrase called beta-fructofuranosidase is not affected ...
... -- Researchers from Weill Cornell Medical College may have ... of the world,s most common blood disorders, beta-thalassemia, commonly ... anemia is the most commonly inherited disease in the ... out of every 100,000 African-Americans. The World Health Organization ...
Cached Biology News:Research suggests parts of UK could be too hot for wine making by 2080 2Avoiding Spleen Removal for Cooley's Anemia SufferersAvoiding spleen removal for Cooley's anemia suf 2Avoiding Spleen Removal for Cooley's Anemia SufferersAvoiding spleen removal for Cooley's anemia suf 3Avoiding Spleen Removal for Cooley's Anemia SufferersAvoiding spleen removal for Cooley's anemia suf 4
Kit includes your choice of reagents for labeling 3 x 10 9 cells and EasySep magnet....
... Complete kit for stabilizing 1.5mg of ... been optimised for the stabilization of aqueous ... traditional methods such as guanidine, phenol/chloroform, silica, ... Our StabMRT Kits contain all the necessary ...
Blasticidin resistance gene (bsr), provided in a pORF plasmid. pORF is an expression plasmid selectable with Ampicillin....
Plasmid expressing Zeocin resistance gene....
Biology Products: