[Protocol] Cell-Type Specific Transfection Protocols (X-treme)
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Cell-Type Specific Transfection Protocols
Experiments below show high level gene expression in mammalian and insect cells transfected using X-tremeGENE Transfection Reagents.
Fast straightforward transfection protocols can be obtained by clicking the cell type below. These protocols are guidelines. Optimal transfection conditions must be determined empirically for your cell culture conditions.
- X-tremeGENE HP DNA Transfection Reagent
- X-tremeGENE 9 DNA Transfection Reagent
- X-tremeGENE siRNA Transfection Reagent
Protocols and results provided by customers
Protocols shown here are provided by customers and have not been verified by Roche scientists. These protocols are guidelines. Optimal transfection conditions must be determined empirically for your cell culture conditions.
X-tremeGENE HP DNA Transfection Reagent
Cell name |
Cell type |
Transfected reporter gene |
Customer protocol(s) |
293 (ATCC® CRL-1573™) |
Human embryonic kidney cell line |
GFP / EGFP-CB2 / Not indicated |
|
293 (ATCC® CRL-1573™) |
Human embryonic kidney cell line |
Stable transfection |
|
Human embryonic kidney cell line |
GFP |
- | |
Human embryonic kidney cell line subclone |
Luciferase |
- | |
293FT |
Human embryonic kidney cell line subclone |
GFP |
|
293T |
Human embryonic kidney cell line expressing SV40 large T antigen |
GFP / Interleukin / Fc fusion protein |
|
3T3 (ATCC®CRL-1658™) |
Mouse embryonic fibroblasts |
GFP |
|
3T3-L1 (ATCC®CRL-173™) |
Mouse embryonic fibroblasts |
SEAP |
- |
A549 (ATCC®CCL-185™) |
Human carcinomic alveolar basal epithelial cells |
SEAP and EGFP |
|
AGS (ATCC®CRL-1739™) |
Human epithelial adenocarcinoma cells |
GFP |
|
C2C12 (ATCC® CRL-1772™) |
Mouse myoblast cell line |
SEAP and EGFP |
|
CHO/dhFr- (ATCC® CRL-9096™) |
Chinese hamster ovary cell line subclone |
Secretable protein |
|
CHO-K1 (ATCC® CCL-61™) |
Chinese hamster ovary cell line subclone |
GFP |
|
Chinese hamster ovary cell line subclone |
Luciferase |
- | |
COS-7 (ATCC® CRL-1651™) |
African green monkey kidney fibroblast-like cell line |
GFP |
|
Dental papilla mesenchymal cells |
Primary cells from the first lower molar of E18.5 Swiss mice |
GFP |
|
EC109 |
Esophageal squamous carcinoma cell line |
Not indicated |
|
EC9706 |
Esophageal squamous carcinoma cell line |
Not indicated |
|
F9 (ATCC® CRL-1720™) |
Mouse embryonal carcinoma cells |
EGFP / GFP |
|
Human primary fibroblasts |
GFP |
- | |
Human embryonic stem cells |
GFP |
- | |
HCT 116 (ATCC® CCL-™) |
Human colorectal carcinoma cell line |
Luciferase |
- |
HeLa (ATCC® CCL-2™) |
Human cervix carcinoma cell line |
GFP |
- |
HepG2 (ATCC® HB-8065™) |
Human hepatoma cell line |
GFP |
|
HepG2.2.15 |
Human hepatoma cell line expressing complete HBV genome |
GFP |
|
Insect cell line (Invitrogen) |
ß-gal |
- | |
HT-29 (ATCC® HTB-38™) |
Human colorectal adenocarcinoma cell line |
GFP |
- |
HT-1080 (ATCC® CCL 121™) |
Human fibrosarcoma cell line |
GFP |
|
HuH-7 (ATCC® JCRB-0403) |
Human hepatoma cell line |
GFP |
- |
HuH-7.5 |
Human hepatoma cell line |
EGFP |
|
Human lymphoblast cell line |
GFP |
- | |
K-562 (ATCC® CCL 243™) |
Human immortalised myelogenous leukemia cell line |
GFP |
- |
L02 |
Immortal hepatic cell line |
RNAi plasmid encoding GFP |
|
LNCaP (ATCC® CRL-1740™) |
Human prostate cancer cell line |
GFP |
|
MC3T3-E1 (ATCC® CRL-2593™) |
Mouse osteoblast-like cells |
sheep C12orf29 protein |
- |
MCF7 (ATCC® HTB-22™) |
Human adenocarcinoma cell line |
GFP |
- |
MDA-MB-231 (ATCC® HTB-26™) |
Human adenocarcinoma cell line |
SEAP |
- |
Primary mouse embryonic fibroblasts |
GFP |
- | |
Human lung fibroblast cells |
GFP |
- | |
PC-3 (ATCC® CRL-1435™) |
Human prostate adenocarcinoma cell line |
GFP |
- |
PC-12 (ATCC® CRL-1721™) |
Rat adrenal pheochromocytoma cell line |
Luciferase |
- |
Plat-E |
Human kidney packaging cell line |
GFP |
|
Porcine ovarian granulosa cells |
Primary cells |
EGFP |
|
PK-15 (ATCC®CCL-33™) |
Porcine kidney epithelial cell line |
GFP |
|
RAW 264.7 (ATCC® TIB-71™) |
Mouse monocyte macrophage cell line |
GFP |
- |
Epithelial Drosophila melanogaster cell line |
GFP |
- | |
Sf9 (ATCC® TIB-71™) |
Epithelial fall armyworm (Spodoptera frugiperda) cells |
ß-gal |
- |
Human mesenchymal stem cells |
GFP |
- | |
Stems cells, mesenchymal |
Murine mesenchymal stem cells (RM26/2) |
Far-red fluorescent protein |
|
Stems cells, mesenchymal |
Murine mesenchymal stem cells (EK8) |
Far-red fluorescent protein |
|
Stems cells, mesenchymal |
Murine mesenchymal stem cells (p53-/-) |
Far-red fluorescent protein |
|
U-2 OS (ATCC® HTB-96™) |
Human osteosarcoma cell line |
GFP |
|
U-87 MG (ATCC® HTB-14™) |
Human epithelial glioblastoma cells |
GFP |
For further information on cells transfected with X-tremeGENE HP DNA Transfection Reagent you can also browse the Transfection Database.
X-tremeGENE 9 DNA Transfection Reagent
Cell name |
Cell type |
Transfected reporter gene |
Customer protocol(s) |
293-A |
Human embryonic kidney cell line |
EGFP |
|
293FT |
Human embryonic kidney cell line subclone |
Luciferase |
|
293T |
Human embryonic kidney cell line |
Luciferase / EGFP / Flag-tagged protein |
|
BF-2 (ATCC® CCL-91™) |
Human embryonic kidney cell line |
RFP and EGFP |
|
C2C12 (ATCC® CRL-1772™) |
Mouse myoblast cell line |
EGFP |
|
CHO-K1 (ATCC® CCL-61™) |
Chinese hamster ovary cell line subclone |
GFP |
- |
CHSE-214 (ATCC® CRL-1681™) |
Chinook salmon embryo (fish cell line) |
EGFP |
|
COS-7 (ATCC® CRL-1651™) |
African green monkey kidney fibroblast-like cell line |
GFP |
|
EPC (ATCC® CRL-2872™) |
Fish cell line (Epithelioma papulosum cyprini) |
RFP |
|
ES-E14TG2a (ATCC® CRL-8121™) |
Mouse embyonic stem cells |
GFP |
- |
GF |
Fish cell line (grouper fin) |
RFP and EGFP |
|
HCT 116 (ATCC® CCL-™) |
Human colorectal carcinoma cell line |
Luciferase |
- |
HeLa (ATCC® CCL-2™) |
Human cervix carcinoma cell line |
EGFP-actin |
|
HeLa (ATCC® CCL-2™) |
Human cervix carcinoma cell line |
GFP |
- |
HepG2 (ATCC® HB-8065™) |
Human hepatoma cell line |
GFP |
- |
HepG2 (ATCC® HB-8065™) |
Human hepatoma cell line |
14 kb plasmid (incl. eGFP; stable) |
|
HINAE |
Japanese flounder (Hirame Natural Embryo) fish cell line |
RFP and EGFP |
|
HT-1080 (ATCC® CCL-121™) |
Human fibrosarcoma cell line |
GFP |
- |
HuH-7 (ATCC® JCRB-0403) |
Human hepatoma cell line |
Luciferase |
|
MCF7 (ATCC® HTB-22™) |
Human adenocarcinoma cell line |
GFP |
- |
Neuro-2a (ATCC® CCL-131™) |
Mouse neuroblastoma cells |
GFP-mortalin fusion protein |
|
NIH-3T3 (ATCC® CRL-1658™) |
Mouse embryonic cell line (fibroblast) |
GFP |
- |
PC-3 (ATCC® CRL-1435™) |
Human prostate adenocarcinoma cell line |
GFP |
- |
PC-12 (ATCC® CRL-1721™) |
Rat adrenal pheochromocytoma cell line |
Luciferase |
- |
Primary pulmonary fibroblasts |
Human pulmonary fibroblasts (primary) |
Not indicated |
|
Porcine granulosa cells |
Porcine granulosa cells (subculture of primary cells) |
GFP fusion protein |
|
Sf9 (ATCC® TIB-71™) |
Epithelial fall armyworm (Spodoptera frugiperda) cells |
GFP |
For further information on cells transfected with X-tremeGENE 9 DNA Transfection Reagent you can also browse the Transfection Database.
X-tremeGENE siRNA Transfection Reagent
Cell Type |
X-tremeGENE |
siRNA1 |
Culture Dish |
Total Volume |
CHO-K1 (ATCC® CCL-61™) |
10 µl |
2 µg |
24-well plate |
550 µl |
COS-7 (ATCC® CRL-1651™) |
2.5 µl |
0.5 µg |
24-well plate |
550 µl |
HEK-293 (ATCC® CRL-1573™) |
5 µl |
0.5 µg |
24-well plate |
550 µl |
HeLa (ATCC® CCL-2™) |
5 µl |
0.5 µg |
24-well plate |
550 µl |
Hep G2 (ATCC® HB-8065™) |
10 µl |
2 µg |
24-well plate |
550 µl |
HT-29 (ATCC® HTB-38™) |
10 µl |
2 µg |
24-well plate |
550 µl |
HTC-116 (ATCC® CCL-247™) |
2.5 µl |
0.5 µg |
24-well plate |
550 µl |
Huh-7 clone 9B |
10 µl |
1 µg |
6-well plate |
1000 µl2 |
MCF7 (ATCC® HTB-22™) |
2.5 µl |
0.5 µg |
24-well plate |
550 µl |
NIH 3T3 (ATCC® CRL-1658™) |
5 µl |
0.5 µg |
24-well plate |
550 µl |
PC-3 (ATCC® CRL-1435™) |
5 µl |
0.5 µg |
24-well plate |
550 µl |
PC-12 (ATCC® CRL-1721™) |
2.5 µl |
0.5 µg |
24-well plate |
550 µl |
10.5 µg corresponds to 67.5 nM siRNA
2medium changed 5 hours after transfection
For further information on cells transfected with X-tremeGENE siRNA Transfection Reagent you can also browse the Transfection Database.
Protocols and results provided by customers
Protocols shown here are provided by customers and can be obtained by clicking the cell type below. These protocols have not been verified by Roche scientists. These protocols are guidelines. Optimal transfection conditions must be determined empirically for your cell culture conditions.
Note: Data and experimental conditions included in "Customer transfection protocols" are the sole responsibility of the customers that have submitted them. Roche was neither involved in establishing the experimental conditions nor in defining the criteria for the performance of the specific assays. Roche therefore cannot take any responsibility for performance or interpretation of results obtained for the biological target parameter(s) described by the authors or other users using a similar experimental approach.
X-tremeGENE HP DNA Transfection Reagent
Cell name |
Cell type |
Transfected reporter gene |
293 (ATCC® CRL-1573™) |
Human embryonic kidney cell line |
Not indicated |
293 (ATCC® CRL-1573™) |
Human embryonic kidney cell line |
GFP |
293 (ATCC® CRL-1573™) |
Human embryonic kidney cell line |
EGFP-CB2 |
293 (ATCC® CRL-1573™) |
Human embryonic kidney cell line |
Stable transfection |
Human embryonic kidney cell line |
EGFP fusion protein | |
Human embryonic kidney cell line subclone |
GFP | |
Human embryonic kidney cell line subclone |
Luciferase | |
Human embryonic kidney cell line subclone |
Pelota/Ubc9 | |
Human embryonic kidney cell line expressing SV40 large T antigen |
GFP | |
Human embryonic kidney cell line expressing SV40 large T antigen |
Interleukin /Fc fusion protein | |
3T3 (ATCC®CRL-1658™) |
Mouse embryonic fibroblasts |
GFP |
A549 (ATCC®CCL-185™) |
Human carcinomic alveolar basal epithelial cells |
EGFP |
AGS (ATCC®CRL-1739™) |
Human epithelial adenocarcinoma cells |
GFP |
C2C12 (ATCC®CRL-1772™) |
Mouse myoblast cell line |
EGFP |
CHO/dhFr- (ATCC® CRL-9096™) |
Chinese hamster ovary cell line subclone |
Secretable protein |
CHO-K1 (ATCC® CCL-61™) |
Chinese hamster ovary cell line subclone |
GFP |
COS-7 (ATCC® CRL-1651™) |
African green monkey kidney fibroblast-like cell line |
GFP |
Primary cells from the first lower molar of E18.5 Swiss mice |
GFP | |
Esophageal squamous carcinoma cell line |
Not indicated | |
Esophageal squamous carcinoma cell line |
Not indicated | |
F9 (ATCC® CRL-1720™) |
Mouse embryonal carcinoma cells |
EGFP |
F9 (ATCC® CRL-1720™) |
Mouse embryonal carcinoma cells |
GFP |
F9 (ATCC® CRL-1720™) |
Mouse embryonal carcinoma cells |
GFP |
HCT 116 (ATCC® CCL-™) |
Human colorectal carcinoma cell line |
Not indicated |
HeLa (ATCC® CCL-2™) |
Human cervix carcinoma cell line |
EGFP fusion protein |
HepG2 (ATCC® HB-8065™) |
Human hepatoma cell line |
GFP |
HepG2 (ATCC® HB-8065™) |
Human hepatoma cell line |
GFP |
HepG2 (ATCC® HB-8065™) |
Human hepatoma cell line |
GFP |
HepG2 (ATCC® HB-8065™) |
Human hepatoma cell line |
GFP and shRNA |
HepG2 (ATCC® HB-8065™) |
Human hepatoma cell line |
HBV genome |
Human hepatoma cell line expressing complete HBV genome |
GFP | |
HT-1080 (stably transfected) |
Human fibrosarcoma cell line |
EGFP-120 kDa nuclear protein |
HuH-7 (ATCC® JCRB-0403) |
Human hepatoma cell line |
Hepatitis B surface antigen |
HuH-7 (ATCC® JCRB-0403) |
Human hepatoma cell line |
Luciferase |
Human hepatoma cell line |
EGFP | |
Immortal hepatic cell line |
RNAi plasmid encoding GFP | |
LNCaP (ATCC® CRL-1740™) |
Human prostate cancer cell line |
GFP |
MCF7 (ATCC® HTB-22™) |
Human adenocarcinoma cell line |
HA-tagged protein |
MCF7 (ATCC® HTB-22™) |
Human adenocarcinoma cell line |
Not indicated |
MEF (ATCC® HTB-22™) |
Primary mouse embryonic fibroblasts |
Not indicated |
Human kidney packaging cell line |
GFP | |
PK-15 (ATCC®CCL-33™) |
Porcine kidney epithelial cell line |
GFP |
Primary cells |
EGFP | |
Murine mesenchymal stem cells (RM26/2) |
Far-red fluorescent protein | |
Murine mesenchymal stem cells (EK8) |
Far-red fluorescent protein | |
Murine mesenchymal stem cells (p53-/-) |
Far-red fluorescent protein | |
U-2 OS (ATCC® HTB-96™) |
Human osteosarcoma cell line |
GFP |
U-87 MG (ATCC® HTB-14™) |
Human epithelial glioblastoma cells |
GFP |
For further information on cells transfected with X-tremeGENE HP DNA Transfection Reagent you can also browse the Transfection Database.
X-tremeGENE 9 DNA Transfection Reagent
Cell name |
Cell type |
Transfected reporter gene |
Human embryonic kidney cell line |
EGFP | |
Human embryonic kidney cell line subclone |
Luciferase | |
Human embryonic kidney cell line |
Luciferase | |
Human embryonic kidney cell line |
EGFP | |
Human embryonic kidney cell line |
Flag-tagged protein | |
BF-2 (ATCC® CCL-91™) |
Bluegill fry (fish cell line) |
RFP and EGFP |
C2C12 (ATCC® CRL-1772™) |
Mouse myoblast cell line |
EGFP |
CHSE-214 (ATCC® CRL-1681™) |
Chinook salmon embryo (fish cell line) |
EGFP |
COS-7 (ATCC® CRL-1651™) |
African green monkey kidney fibroblast-like cell line |
EGFP-fusion protein |
COS-7 (ATCC® CRL-1651™) |
African green monkey kidney fibroblast-like cell line |
GFP |
EPC (ATCC® CRL-2872™) |
Fish cell line (Epithelioma papulosum cyprini) |
RFP |
Fish cell line (grouper fin) |
RFP and EGFP | |
HeLa (ATCC® CCL-2™) |
Human cervix carcinoma cell line |
EGFP-actin |
HeLa (ATCC® CCL-2™) |
Human cervix carcinoma cell line |
GFP |
HepG2 (ATCC® HB-8065™) |
Human hepatoma cell line |
14 kb plasmid (incl. eGFP; stable) |
Japanese flounder (Hirame Natural Embryo) fish cell line |
RFP and EGFP | |
HuH-7 (ATCC® JCRB-0403) |
Human hepatoma cell line |
Luciferase |
MCF7 (ATCC® HTB-22™) |
Human adenocarcinoma cell line |
FAK-enconding shRNA plasmid |
Neuro-2a (ATCC® CCL-131™) |
Mouse neuroblastoma cells |
GFP-mortalin fusion protein |
PC-12 (ATCC® CRL-1721™) |
Rat adrenal pheochromocytoma cell line |
EGFP fusion protein |
Human pulmonary fibroblasts (primary) |
Not indicated | |
Porcine granulosa cells (subculture of primary cells) |
GFP fusion protein | |
Sf9 (ATCC® TIB-71™) |
Epithelial fall armyworm (Spodoptera frugiperda) cells |
GFP |
For further information on cells transfected with X-tremeGENE 9 DNA Transfection Reagent you can also browse the Transfection Database.