Name | Catalog | Size |
INVI DNA Transfection Reagent™ | IV1214025 | 0.25 ml |
INVI DNA Transfection Reagent™ | IV1214050 | 0.5 ml |
INVI DNA Transfection Reagent™ | IV1214075 | 0.75 ml |
INVI DNA Transfection Reagent™ | IV1214100 | 1 ml |
INVI DNA Transfection Reagent™ | IV1214150 | 1.5 ml |
INVI DNA Transfection Reagent™ | IV1214300 | 3 ml |
INVI DNA Transfection Reagent™
CatNo:IV1214
1.Description
INVI DNA Transfection Reagent™ is a newly developed reagent for the transfection of DNA into eukaryotic cells. Advantages:
-The highest transfection efficiency in many cell types.
-DNA-INVI DNA Transfection Reagent™complexes can be directly added to cells in culture medium.
-It is not necessary to remove DNA-INVI DNA Transfection Reagent™ complexes following transfection.
-The complexes can be removed after 4-6 hours by replacing with refresh medium (optional)
2.Contents and Storage
Contents: INVI DNA Transfection Reagent™ is supplied in liquid form .
Storage : Store at 4oC. DO NOT FREEZE.
3.Transfection Procedure(24-Well Format)
A. For each transfection sample,prepare DNA-INVI DNA Transfection Reagent™ complexes as follows:
a. Dilute DNA in 50 µl of Opti-MEM. Mix gently.
b. Mix INVI DNA Transfection Reagent™ gently before use,then dilute the appropriate amount in 50 µl of Opti-MEM. Mix gently and incubate for 5 minutes at room temperature.
c. After the 5 minute incubation,combine the diluted DNA with the diluted INVI DNA Transfection Reagent™(total volume is 100 µl). Mix gently and incubate for 20 minutes at room temperature to allow the DNA-INVI DNA Transfection Reagent™ complexes to form. The solution may appear cloudy,but this will not inhibit the transfection.
Note: DNA-INVI DNA Transfection Reagent™ complexes are stable for at least 5 hours at room temperature.
B. Add the 100 µl of DNA-INVI DNA Transfection Reagent™ complexes to each well.
C. Incubate the cells at 37oC in a CO2 incubator for 24-48 hours until they are ready to assay for transgene expression. It is not necessary to remove the complexes or change the medium; however,growth medium may be replaced after 4-6 hours without loss of transfection activity.
D. The table of Transfection Procedure:
Culture Vessel | Surface Area per Well (cm2) | Volume of Plating Medium | DNA (µg) and Dilution Volume (µl) | INVI DNA Transfection Reagent™ (µl) and Dilution Volume (µl) |
96-well | 0.3 | 100 µl | 0.2 µg in 25 µl | 0.5 µl in 25 µl |
24-well | 2 | 500 µl | 0.8 µg in 50 µl | 2.0 µl in 50 µl |
12-well | 4 | 1 ml | 1.6 µg in 100 µl | 4.0 µl in 100 µl |
35-mm | 10 | 2 ml | 4.0 µg in 250 µl | 10 µl in 250 µl |
6-well | 10 | 2 ml | 4.0 µg in 250 µl | 10 µl in 250 µl |
60-mm | 20 | 5 ml | 8.0 µg in 0.5 ml | 20 µl in 0.5 ml |
10-cm | 60 | 15 ml | 24 µg in 1.5 ml | 60 µl in 1.5 ml |
4.Important Guidelines
Follow these guidelines when performing transfections:
A.The ratio of DNA (in µg) : INVI DNA Transfection Reagent™ (in µl) to use when preparing complexes should be 1:1 to 1:4for most cell lines.
B. It is CRITICAL to transfect cells at high cell density. 70-90% confluence the time of transfection is recommended to obtain high efficiency and expression levels and to minimize decreased cell growth associated with high transfection activity. Lower cell densities are suitable with optimization of conditions.
C. DO NOT add antibiotics to media during transfection as this will cause cell death.
5.Optimizing Transfection
To obtain the highest transfection efficiency and low non-specific effects,optimize transfection conditions by varying DNA and INVI DNA Transfection Reagent™ concentrations, and cell number. Make sure that cells are greater than 90% confluent and vary DNA(µg)/INVI DNA Transfection Reagent™ (µl) ratios from 1/0.5 to 1/5.