ScriptCap™ Cap 1 Capping System C-SCCS1710
产品简介:北京中北林格供应ScriptCap™ Cap 1 Capping System C-SCCS1710,中北林格是cellscript总代理,提供cellscript产品货号报价查询。其他品牌包括LGC Biosearch Technologies总代理,epicentre总代理,lucigen总代理,illumina总代理,ICLLAB总代理,ImmunoReagents总代理。
ScriptCap™ Cap 1 Capping System C-SCCS1710 品牌介绍:
CELLSCRIPT专注于再生医学,细胞再生学以及细胞疗法。细胞、组织或器官的移植ZL及修复中应用的细胞或生物体。CellScript还生产用于传递和翻译的合成修饰mRNA。
ScriptCap™ Cap 1 Capping System C-SCCS1710 规格详情名称:ScriptCap™ Cap 1 Capping System
分类:Post-Transcriptional Capping货号:C-SCCS1710规格:10 reactions产品详情:For post-transcriptional capping of in vitro transcribed RNA with ScriptCap Capping Enzyme, ScriptCap 2'-O-Methyltransferase, GTP and SAM. Either Cap 0 or Cap 1 caps can be built with efficiencies approaching 100%. This cannot be accomplished with co-transcriptional capping methods.
ScriptCap™ Cap 1 Capping System C-SCCS1710 产品详情
A. Synthesis of Cap 0 RNA from Uncapped RNA 1. The protocol below was designed to build a cap 1 structure on 5' end of 50-60 µg of uncapped RNA. If a cap 0 structure is desired, replace the ScriptCap 2'-O-Methyltransferase in Step 3 with an equivalent volume of RNase-Free Water. Combine the following reagents:
Standard ScriptCap Cap 1 Capping Reaction (step 1) | |
Component | Amount |
RNase-Free Water | x µl |
In vitro transcribed RNA, 50-60 µg | ≤67 µ |
Total Volume | 67 µl |
2. Incubate at 65o C for 5-10 minutes, then transfer to ice. 3. While the heat-denatured RNA is coo领 on ice, prepare a “Cocktail” of the following reaction components together in a separate tube.
Standard ScriptCap Cap 1 Capping Reaction (step 3) | |
Component | Amount |
10X ScriptCap Capping Buffer | 10 µl |
10 mM GTP | 10 µl |
20 mM SAM | 2.5 µl |
ScriptGuard RNase Inhibitor | 2.5 µl |
ScriptCap 2'-O-Methyltransferase (100 U/µl) | 4 µl |
Total Volume | 29 µl |
4. Just prior to starting the reaction, add the ScriptCap Capping Enzyme to the Cocktail from Step 3 and then combine this with the uncapped RNA solution from Step 1.
Standard ScriptCap Cap 1 Capping Reaction (step 4) | |
Component | Amount |
Cocktailed reaction components (from step 3) | 29 µl |
ScriptCap Capping Enzyme (10 U/µl) | 4 µl |
Heat-denatured RNA (from step 1) | 67 µl |
Total Reaction Volume | 100 µl |
5. Incubate at 37o C for 30 minutes. 6. The Capped RNA can now be purified, or it can be 3' polyadenylated without purification by adding the reaction mixture directly to an A-Plus Poly(A) Polymerase Tai领 Kit reaction (sold separately).
ScriptCap™ Cap 1 Capping System Contents (10 Reactions) | |
Kit Component | Reagent Volume |
ScriptCap™ Capping Enzyme, 10 U/µl (red cap) | 40 µl |
ScriptCap™ 2'-O-Methyltransferase, 100 U/µl (blue cap) | 40 µl |
10X ScriptCap™ Capping Buffer. 0.5 M Tris-HCl, pH 8.0, 60 mM KCl and 12.5 mM MgCl2 | 100 µl |
10 mM GTP Solution | 100 µl |
20 mM S-adenosyl-methionine (SAM) | 25 µl |
ScriptGuard™ RNase Inhibitor, 40 U/µl | 25 µl |
RNase-Free Water | 1.4 ml |
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ScriptCap™ Cap 1 Capping System C-SCCS1710 关键词:
ScriptCap™ Cap 1 Capping System
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