Navigation Links
Uses and Applications of FuGENE 6 Transfection Reagent

Specific technique considerations

Co-transfections

First, verify that both of your DNA constructs work in your cell line. Also, determine the ratio of DNA to FuGENE 6 Reagent (for example, 1 g of DNA and 3 l FuGENE 6 Reagent) that provides good expression levels. Then, perform the transfection, ensuring the DNA / FuGENE 6 Reagent ratio takes into account the total amount of DNA from the two constructs. For example, in the above experiment the co-transfection of 1 g each of two carefully measured DNA constructs would require 6 l of FuGENE 6 Transfection Reagent (2 g DNA to 6 l of FuGENE 6 Transfection Reagent). FuGENE 6 Transfection Reagent is very gentle, so there will not be a cytotoxic effect when an excess of FuGENE 6 Transfection Reagent is used.

Using spinner flasks

The special components (e.g., Pluronic F-68) that are added to the spinner flask media for cell protection can interfere with transfection. To overcome this problem:

At the time of subculturing, resuspend the cells in medium that does not contain protective reagents, then perform the transfection in a flask or Petri dish for one hour as suggested in the package insert. For example, for 1 g DNA, use 3 l FuGENE 6 Reagent for each 100,000-200,000 cells. The cell concentration may be several times the concentration you will use for seeding. After allowing an hour for transfection, dilute into your normal suspension medium, then place it into the spinner flask.

Producing stable transfectants

Do not use a large number of plates, or plate fewer cells per plate. FuGENE 6 Reagent is so gentle and efficient that you will obta in many transfected cells. Use a good construct and good selective medium, and you will obtain a large number of clones.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. A Versatile Power Supply for All Electrophoresis Applications
2. Highest Possible Transformation Efficiencies for High-Throughput Applications
3. Unique DNA Polymerase Formulation Excels in a Broad Range of PCR Applications
4. High-Throughput System Generates Ultra-Pure PCR Products Suitable for All Applications
5. Optimizing Büchi® Rotary Evaporator Applications
6. Optimizing Büchi® Rotary Evaporator Applications
7. Optimizing Bchi Rotary Evaporator Applications
8. Uses and Applications of X-tremeGENE Q2 Transfection Reagent
9. Uses and Applications of FuGENE 6 Transfection Reagent
10. Uses and Applications of FuGENE 6 Transfection Reagent
11. Uses and Applications of FuGENE 6 Transfection Reagent
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:6/23/2016)... ... June 23, 2016 , ... ... announce the launch of their brand, UP4™ Probiotics, into Target stores nationwide. The ... is proud to add Target to its list of well-respected retailers. This list ...
(Date:6/23/2016)... Md. , June 23, 2016 A person ... from the crime scene to track the criminal down. ... the U.S. Food and Drug Administration (FDA) uses DNA evidence ... Sound far-fetched? It,s not. The FDA ... sequencing to support investigations of foodborne illnesses. Put as simply ...
(Date:6/23/2016)... -- The Prostate Cancer Foundation (PCF) is pleased to announce 24 new ... prostate cancer. Members of the Class of 2016 were selected from a pool ... Read More About the Class of 2016 PCF Young Investigators ... ... ...
(Date:6/23/2016)... ... June 23, 2016 , ... ... the launch of the Supplyframe Design Lab . Located in Pasadena, Calif., ... the future of how hardware projects are designed, built and brought to market. ...
Breaking Biology Technology:
(Date:6/20/2016)... , June 20, 2016 Securus ... justice technology solutions for public safety, investigation, corrections ... the prisons involved, it has secured the final ... (DOC) facilities for Managed Access Systems (MAS) installed. ... additional facilities to be installed by October, 2016. ...
(Date:6/7/2016)... 7, 2016  Syngrafii Inc. and San Antonio ... that includes integrating Syngrafii,s patented LongPen™ eSignature "Wet" ... collaboration will result in greater convenience for SACU ... while maintaining existing document workflow and compliance requirements. ... Highlights: ...
(Date:6/1/2016)... June 1, 2016 Favorable Government ... Administration and Criminal Identification to Boost Global Biometrics System ... released TechSci Research report, " Global Biometrics Market ... Forecast and Opportunities, 2011 - 2021", the global biometrics ... 2021, on account of growing security concerns across various ...
Breaking Biology News(10 mins):