中文名稱：脂質體型DNA高效轉染試劑

英文名稱： Lipopro DNAHitrans Reagent

目錄號：QYR024A, QYR024B

規格：0.8ml, 1.5ml

保存條件：4℃

保質期：1年

產品描述：

DNA高效轉染試劑用于真核細胞轉染，包括體外轉染與體內轉染。與其他轉染方法，例如磷酸鈣共沉淀、顯微注射、基因槍、DEAE-dextran轉染、脂質體轉染等相比，DNA高效轉染試劑實驗操作簡單、轉染效率高、細胞毒性低、價格便宜。本試劑也可用于實驗動物體內轉染。通過靜脈、心室、皮下、氣管、腹腔注射等方式，實現包括小鼠、大鼠、蝌蚪及鴨子等多種實驗動物的體內轉染。

本試劑不可用于siRNA轉染。如需要做siRNA轉染或DNA/siRNA共轉染實驗，請使用EasyTrans Reagent (QYR026)。

體外轉染實驗流程：

以6孔板為例。如需放大或縮小體系，參見表格1。

1. 貼壁細胞：實驗前一天在6孔板中接種細胞。轉染前，用PBS洗細胞2次，加入2ml基礎培養基（不含血清、抗生素）。懸浮細胞：轉染前將細胞離心，使用2ml基礎培養基將細胞重懸。
2. 將4ug DNA加入250ul PBS中，吹打混勻。
3. 將10ul DNA高效轉染試劑加入250ul PBS中，吹打混勻，靜置5min。
4. 將稀釋的DNA加入稀釋的DNA高效轉染試劑中，吹打混勻，靜置20min。
5. 將上述轉染混合物500ul加入細胞中。輕輕前后左右晃動6孔板，使轉染混合物與細胞充分接觸。
6. 將細胞放回細胞培養箱，繼續培養4-6h，之后將培養基換成完全培養基（含血清、抗生素）。

體內轉染實驗流程：

1. 將10ug DNA 加入50ul PBS 中，吹打混勻。
2. 將5ul DNA高效轉染試劑加入50ul PBS中，吹打混勻，靜置5min。
3. 將稀釋的DNA高效轉染試劑加入稀釋的DNA中，吹打混勻，靜置20min。
4. 注射動物。詳見表格2。

Description

DNAHitrans is a superior proprietary formulation optimized for the transfection of DNA into many eukaryotic cells with ease of use, maximal transfection efficiency and minimal cytotoxicity to a wide variety of other transfection techniques including calcium phosphate coprecipitation, electroporation, microinjection, biolistic particle delivery, complex formation with DEAE-dextran and lipofectamine-mediated DNA transfection. In addtion, because of its high transfection efficiency, no or minimal toxicity and immunogenicity, DNAHitrans also is a superior alternative for the transfection of DNA into many animals in vivo including mice, rats, tadpoles and ducks via intravenous, intraventricular, subcutaneous, tracheal and intraperitoneal injections.

in vitro transfection

Use the following procedure to transfect mammalian cells in a 6-well format. For other formats, please see table 1.

1. Adherent cells: Cells should be seeded a day prior to transfection in 6-well plates at an appropriate density. Before transfection, cells should be washed twice with PBS, then cultured in 2 mL of basal medium (containing no additives ie serum, antibiotics or other proteins).

Suspension cells: Just prior to preparing complexes, plate cells at an appropriate density in 2 mL of basal medium (containing no additives ie serum, antibiotics or other proteins).

2. Dilute 4 μg DNA in 250 μL PBS buffer and mix by pipetting up-down.
3. Dilute 10 μL DNAHitrans in 250 μL PBS, mix by pipetting up-down gently and incubate for 5 min at room temperature. Then combine the diluted DNA with diluted DNAHitrans, mix immediately by pipetting up-down and incubate for 20 minutes at room temperature.
4. Add the 500 μL of mixture to each well dropwise. Mix gently by rocking the plate back and forth.
5. Incubate cells at 37℃ in a CO₂ incubator for 4-6 hours and then the medium is replaced by complete medium.

in vivo Transfection

1. Dilute 10 μg of DNA in 50 μL of sterile PBS solution. Vortex gently and spin down briefly.
2. Dilute 5 μL of DNAHitrans in 50 μL of sterile PBS solution. Vortex gently and incubate for 5 min at room temperature.
3. Add the diluted 50 μL of DNAHitrans to the diluted 50 μL of DNA. Mix immediately by pipetting up-down immediately and spin down briefly. Incubate 20 min at room temperature.
4. Inject animals. Please see table 2.

Table 1. Scaling Up or Down Transfections

Table 2. Suggested Amount of DNA and Maximum Injection Volume

* DNAHitrans reagent is not recommended for siRNA transfection. For Single siRNA transfection or DNA/siRNA co-transfection experiment, another transfection reagent (EasyTrans:QYR026) is recommended.