BLOCK IT U6 ENTRY VECTOR KIT 20 RXNS

产品编号：K494500 品牌：invitrogen 原厂货号：K494500
产品分类：DNA和RNA纯化 > RNA干扰 > RNAi载体（shRNA)
应用分类：暂定分类 > Gateway® 克隆 > 目的载体

包装：

运保温度:

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其它组分：

pENTR./U6 vector,linearized 【包装：4 x 10 μl，，运保温度：-20°C】

10X Oligo Annealing Buffer【包装：250 μl，，运保温度：-20°C】

DNase/RNase-Free Water【包装：3 x 1.5 ml，，运保温度：-20°C】

5X Ligation Buffer【包装：80 μl，，运保温度：-20°C】

T4 DNA Ligase【包装：20 μl，，运保温度：-20°C】

U6 Forward Sequencing Primer【包装：20 μl，，运保温度：-20°C】

M13 Reverse Primer【包装：20 μl，，运保温度：-20°C】

LacZ double-stranded (ds) Control Oligo【包装：4 μl，，运保温度：-20°C】

pcDNA.1.2/V5-GW/lacZ control plasmid【包装：20 μl，，运保温度：-20°】

S.O.C. Medium【包装：6 ml，，运保温度：-80°】

TOP10 cells【包装：21 x 50 μl，，运保温度：-80°】

pUC19 Control DNA【包装：50 μl，，运保温度：-80°】

描述：

The BLOCK-iT™ U6 RNAi Entry Vector provides a simple, streamlined approach to cloning short hairpin RNA (shRNA) sequences for testing in transient transfections for RNA interference (RNAi), a preferred technique for analyzing gene downregulation. An easy cloning process places an ~50-bp oligonucleotide DNA immediately following a U6 pol III type promoter (Figure 1). Expression of this RNAi cassette forms a shRNA molecule in the cell that will be processed and act as short interfering RNA (siRNA) that will generate the RNAi effect.

Delivery of the U6 RNAi Cassette
Once cloning is complete, the U6 Entry Vector is ready to be used immediately in transient transfections using a reagent such as Lipofectamine™ 2000. This makes this system ideal for initial shRNA screenings in many mammalian cell types. As an alternative in hard-to-transfect or non-dividing cell types, or to deliver to animal model systems, the RNAi cassette can easily be recombined into a BLOCK-iT™ Viral RNAi 查看更多vector. For stable shRNA lentiviral delivery and expression, recombine the BLOCK-iT™ U6 Entry Vector with the pLenti6/BLOCK-iT™ RNAi Vector. For transient delivery to challenging cell types using adenoviral transduction, recombine with the pAd/BLOCK-iT™ RNAi Vector.

注意事项：

The BLOCK-iT™ U6 Entry Vector Kit contains two boxes. The cloning box contains four tubes of linearized vector at 5 ng/tube, enough to complete 5 cloning reactions per tube. Also included are 250 µl 10X annealing buffer, 20 µl T4 DNA ligase, 80 µl 5X DNA ligase buffer, 1.4 ng lacZ control oligo, 10 µl lyophilized lacZ control plasmid, 1.5 ml DNase/RNase-free water, and 2 µg each of forward and reverse sequencing primer. Store the vectors, buffers, control oligo and plasmid, water, and sequencing primers at -20°C. The One Shot® box contains transformation reagents including twenty-one 50-µl aliquots of One Shot® TOP10 Chemically Competent E. coli, S.O.C. medium, and a pUC19 supercoiled control plasmid. Store at -80°C. All reagents are guaranteed stable for 6 months when properly stored.