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DNA高效转染试剂 DNAHitrans Reagent  
   目录号 QYR024    
规格
目录价
促销价
库存
购买
0.8ml 0.00 950.00
20
   
1.5ml 0.00 1550.00
10
   
图片描述:
1.5ml
文件与质量管理
COA Datasheet
商品基本信息:

名称:DNA高效转染试剂

英文名称:DNAHitrans Reagent

目录号:QYR024A, QYR024B

规格:0.8ml, 1.5ml

保存条件:4℃

保质期:1年

 

产品描述:

DNA高效转染试剂用于真核细胞转染,包括体外转染与体内转染。与其他转染方法,例如磷酸钙共沉淀、显微注射、基因枪、DEAE-dextran转染、脂质体转染等相比,DNA高效转染试剂实验操作简单、转染效率高、细胞毒性低、价格便宜。本试剂也可用于实验动物体内转染。通过静脉、心室、皮下、气管、腹腔注射等方式,实现包括小鼠、大鼠、蝌蚪及鸭子等多种实验动物的体内转染。

本试剂不可用于siRNA转染。如需要做siRNA转染或DNA/siRNA共转染实验,请使用EasyTrans Reagent (QYR026)。

 

 

体外转染实验流程:

以6孔板为例。如需放大或缩小体系,参见表格1。

  1. 贴壁细胞:实验前一天在6孔板中接种细胞。转染前,用PBS洗细胞2次,加入2ml基础培养基(不含血清、抗生素)。悬浮细胞:转染前将细胞离心,使用2ml基础培养基将细胞重悬。
  2. 将4ug DNA加入250ul PBS中,吹打混匀。
  3. 将10ul DNA高效转染试剂加入250ul PBS中,吹打混匀,静置5min。
  4. 将稀释的DNA加入稀释的DNA高效转染试剂中,吹打混匀,静置20min。
  5. 将上述转染混合物500ul加入细胞中。轻轻前后左右晃动6孔板,使转染混合物与细胞充分接触。
  6. 将细胞放回细胞培养箱,继续培养4-6h,之后将培养基换成完全培养基(含血清、抗生素)。

 

 

 

体内转染实验流程:

  1. 将10ug DNA 加入50ul PBS 中,吹打混匀。
  2. 将5ul DNA高效转染试剂加入50ul PBS中,吹打混匀,静置5min。
  3. 将稀释的DNA高效转染试剂加入稀释的DNA中,吹打混匀,静置20min。
  4. 注射动物。详见表格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).

  1. Dilute 4 μg DNA in 250 μL PBS buffer and mix by pipetting up-down.
  2. 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.
  3. Add the 500 μL of mixture to each well dropwise. Mix gently by rocking the plate back and forth.
  4. 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

Culture vessel

Surface area per well (cm2)

Volume of plating medium

DNA in PBS volume

DNAHitrans in PBS volume

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-well

10

2 mL

4.0 μg in 250 uL

10 μL in 250 μL

6-well

10

2 mL

4.0 μg in 250 uL

10 μL in 250 uL

60-mm

20

5 mL

8.0 μg in 0.5 μL

20 μL in 0.5 mL

10-cm

30

15 mL

24 μg in 1.5 mL

60 μL in 1.5 mL

 

 

Table 2. Suggested Amount of DNA and Maximum Injection Volume

Animal

Route of injection

Suggested amount of DNA (μg)

Maximum injection volume (μL)

Adult mouse

Intravenous

25-125

400-600

Retroorbital

40

200

Intraperitoneal

100

600

Heart

50

200

Lung instillation

20

300

Subcutaneous tumor

10

100

Nude mouse

Intravenous

50

200

Subcutaneous tumor

10

100

 

 

 

 

* 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.

 

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