中文名称：脂质体型DNA高效转(zhuan)染(ran)试剂

英文名称(cheng)： Lipopro 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(miao)述：

DNA高效转(zhuan)染(ran)(ran)试剂(ji)用于(yu)真核(he)细胞转(zhuan)染(ran)(ran)，包(bao)括(kuo)(kuo)体(ti)(ti)外转(zhuan)染(ran)(ran)与体(ti)(ti)内转(zhuan)染(ran)(ran)。与其他(ta)转(zhuan)染(ran)(ran)方(fang)法，例如磷酸(suan)钙共沉淀、显微注(zhu)射、基因(yin)枪(qiang)、DEAE-dextran转(zhuan)染(ran)(ran)、脂质体(ti)(ti)转(zhuan)染(ran)(ran)等(deng)相(xiang)比，DNA高效转(zhuan)染(ran)(ran)试剂(ji)实(shi)(shi)验操(cao)作(zuo)简单、转(zhuan)染(ran)(ran)效率(lv)高、细胞毒(du)性低(di)、价格便(bian)宜。本试剂(ji)也可用于(yu)实(shi)(shi)验动物体(ti)(ti)内转(zhuan)染(ran)(ran)。通过静脉(mai)、心室(shi)、皮(pi)下、气(qi)管、腹腔注(zhu)射等(deng)方(fang)式，实(shi)(shi)现包(bao)括(kuo)(kuo)小鼠、大鼠、蝌蚪及鸭(ya)子等(deng)多种实(shi)(shi)验动物的体(ti)(ti)内转(zhuan)染(ran)(ran)。

本(ben)试(shi)剂不可(ke)用(yong)于(yu)siRNA转(zhuan)染(ran)。如需要做siRNA转(zhuan)染(ran)或(huo)DNA/siRNA共转(zhuan)染(ran)实验，请使用(yong)EasyTrans Reagent (QYR026)。

体外转(zhuan)染(ran)实验流程：

以6孔(kong)板为例。如需(xu)放大(da)或缩小体系，参见表格1。

1. 贴壁细胞：实验前一天在6孔板(ban)中(zhong)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(fu)细(xi)胞(bao)(bao)：转染前(qian)将细(xi)胞(bao)(bao)离(li)心，使用2ml基(ji)础培养(yang)基(ji)将细(xi)胞(bao)(bao)重(zhong)悬。
2. 将4ug DNA加入250ul PBS中，吹(chui)打混匀(yun)。
3. 将(jiang)10ul DNA高(gao)效转染试(shi)剂(ji)加入(ru)250ul PBS中，吹打混匀，静(jing)置(zhi)5min。
4. 将稀(xi)(xi)释(shi)的DNA加入稀(xi)(xi)释(shi)的DNA高效转(zhuan)染试(shi)剂中(zhong)，吹打混匀，静置20min。
5. 将上述转(zhuan)染(ran)混(hun)(hun)合物500ul加(jia)入细胞(bao)中(zhong)。轻(qing)轻(qing)前后左右晃动6孔(kong)板(ban)，使(shi)转(zhuan)染(ran)混(hun)(hun)合物与细胞(bao)充分接触(chu)。
6. 将细(xi)胞(bao)放回细(xi)胞(bao)培养(yang)(yang)箱(xiang)，继续(xu)培养(yang)(yang)4-6h，之后将培养(yang)(yang)基换成完全培养(yang)(yang)基（含血清、抗生素(su)）。

体(ti)内转染实(shi)验流(liu)程：

1. 将10ug DNA 加入(ru)50ul PBS 中(zhong)，吹打混匀。
2. 将5ul DNA高效转(zhuan)染试(shi)剂加入50ul PBS中，吹打(da)混匀，静置5min。
3. 将稀(xi)释的(de)DNA高效(xiao)转染试剂加入稀(xi)释的(de)DNA中，吹打混(hun)匀，静置20min。
4. 注射动物。详见表格(ge)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.