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CRISPR-Cas 9

CRISPR-Cas 9: Reagents for CRISPR-Cas9

Reagents and kits used in CRISPR/Cas 9

T7 mScript™ Standard mRNA Production System

Simply the best way to make in vitro mRNA. Combines the benefits of high-yield transcription with those of post-transcriptional capping and tailing all in one kit.

CellScript

Catalogue No.DescriptionPack SizePriceQty
C-MSC100625T7 mScript™ Standard mRNA Production System25 reactions £731.00 Quantity Add to Order
C-MSC11610T7 mScript™ Standard mRNA Production System10 reactions £334.00 Quantity Add to Order

Description

  • All-in-one kit to produce uridine-containing, fully 5’ capped and 3’ poly(A) tailed mRNA (U-mRNA).
  • Combine the benefits of high-yield transcription with post-transcriptional capping and tailing.
  • Obtain virtually 100% 5’-capped transcripts.
  • Create variable 3’ poly(A) tail lengths, from 50 to >300 A’s.
  • Two kit sizes to produce 0.6 or 1.5 mg of U-mRNA.
  • ScriptGuard™ RNase Inhibitor included to protect synthesized mRNA from RNase degradation.

Product is for research use only (RUO)

Product Description:

The T7 mScript™ Standard mRNA Production System V2 provides all enzymes and reagents for making canonical uridine (U)-containing, 5'-capped and 3'-polyadenylated mRNA. The kit includes reagents for four workflow modules, (1) high-yield in vitro transcription of linear double-stranded DNA templates using the T7 mScript™ Enzyme Solution and the canonical nucleotides ATP, CTP, GTP and UTP, (2) enzymatic capping of the RNA using the ScriptCap™ Cap 1 Capping System (contains both ScriptCap™ Capping Enzyme and 2'-O-Methyltransferase) for making mRNA with a Cap 1 cap structure, (3) A-Plus™ Poly(A) Polymerase for adding a 3' poly(A) tail and (4) 5 M NH4OAc as a convenient RNA/mRNA purification method. ScriptGuard™ RNase Inhibitor is included in the kit to protect synthesized mRNA from RNase degradation.

Post-transfection, capped and tailed mRNA has increased stability and translation efficiency in most eukaryotic cell lines. The mScript™ System improves upon co-transcriptional capping and tailing methods by ensuring virtually 100% transcript capping and enabling users to define their desired tail lengths, even greater than 300 A's. In this way, poly(A) tail lengths can be generated much longer than is possible using a template-encoded tail. mRNAs with longer poly(A) tails are expressed for longer periods of time in cells. mScript™ U-mRNA is suitable for use in transfection and microinjection experiments as well as in vitro translation systems.

For reduction of immunogenicity, combine mScript™ U-mRNA with the Min-Immune™ Gold dsRNA Removal Kit to produce virtually dsRNA-free (to <0.005% of sample [LLOQ]), ultra-low immunogenicity mRNA that is suitable for downstream applications such as cell and gene therapy research and mRNA vaccine development.

CELLSCRIPT™ also offers the INCOGNITO™ T7 mScript™ N1meΨ-mRNA Production System and INCOGNITO™ T7 mScript™ Ψ-mRNA Production System to produce low immunogenicity mRNA containing the modified nucleotides N1-methyl-pseudouridine and pseudouridine, respectively.

Materials Supplied:

Important Store at -20°C in a freezer without a defrost cycle. Do not store at -70°C.

 

 

T7 mScript™ Standard mRNA Production System V2 Kit Contents (Module 1 of 4)
Kit Module Kit Component Reagent Volume
C-MSC11610
10 Reactions
C-MSC100625
25 Reactions
In Vitro Transcription T7 mScript™ Enzyme Solution 20 µl 50 µl
10X T7 mScript™ Transcription Buffer II 20 µl 50 µl
100 mM Dithiothreitol (DTT) 20 µl 50 µl
NTP Solution
25 mM each GTP, ATP, UTP, CTP
80 µl 180 µl
RNase-Free DNase I, 1 U/μl 10 µl 25 µl


T7 mScript™ Standard mRNA Production System V2 Kit Contents (Module 2 of 4)
Kit Module Kit Component Reagent Volume
C-MSC11610
10 Reactions
C-MSC100625
25 Reactions
Post-Transcriptional Capping ScriptCap™ Capping Enzyme, 10 U/μl 40 µl 100 µl
ScriptCap™ 2'-O-Methyltransferase, 100 U/μl 40 µl 100 µl
10X ScriptCap™ Capping Buffer
0.5 M Tris-HCl (pH 8.0), 60 mM KCl and 12.5 mM MgCl2
100 µl 250 µl
20 mM S-adenosyl-methionine (SAM) 50 µl 125 µl
20 mM GTP 50 µl 125 µl


T7 mScript™ Standard mRNA Production System V2 Kit Contents (Module 3 of 4)
Kit Module Kit Component Reagent Volume
C-MSC11610
10 Reactions
C-MSC100625
25 Reactions
Poly(A) Tailing A-Plus™ Poly(A) Polymerase, 4 U/µl 52 µl 130 µl
10X A-Plus™ Poly(A) Tailing Buffer
0.5 M Tris-HCl (pH 8.0), 2.5 M NaCl and 100 mM MgCl2
125 µl 300 µl
20 mM ATP 60 µl 150 µl


T7 mScript™ Standard mRNA Production System V2 Kit Contents (Module 4 of 4)
Kit Module Kit Component Reagent Volume
C-MSC11610
10 Reactions
C-MSC100625
25 Reactions
Common Usage ScriptGuard™ RNase Inhibitor, 40 U/µl 35 µl 90 µl
RNase-Free Water 3 x 1.4 ml 12 ml
5 M Ammonium Acetate 3 x 1.4 ml 12 ml

T7 Control Template DNA: Is a linearized 4.1 kb plasmid that contains a T7 promoter followed by a phage lambda dsDNA insert that encodes a 1,375 base runoff transcript. The Control Template DNA is provided at a concentration of 0.5 µg/µl in T10E1 Buffer (10 mM Tris-HCl, pH 7.5, 1 mM EDTA).

Materials Required, but not Supplied

  • A DNA template for transcription of your RNA of interest
  • Materials or kits for purification of the RNA product
  • RNase-free TE Buffer (10 mM Tris-HCl, pH 7.5, 1 mM EDTA)
  • Optional: TE saturated phenol/chloroform, 0.5-1 M EDTA
 

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Protocols

Protocols for: mScript™ mRNA Production System

 

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T7 mScript™ Standard mRNA Production System

 

Please note: all protocols are the responsibility of the product supplier

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

References

References:

Nicholson, B. L., et al. (2010) Tombusvirus recruitment of host translational machinery via the 3' UTR

Suzuki, T., et al. (2010) Full-term mouse development by abolishing Zn2 -dependent metaphase II arrest without Ca2 release

Palavicini, J. P., et al. (2009) An extra double-stranded RNA binding domain confers high activity to a squid RNA editing enzyme

Jemielity, J. et al., (2003) RNA 9 , 1108.Grudzien, E. et al., (2004) RNA 10 , 1479.Pasquinelli, A.E. et al., (1995) RNA 1 , 957.Stepinski, J. et al., (2001) RNA 7 , 1486.Meis, R. and Meis, J.E . (2006) EPICENTRE Forum 13 (4) 5.Kuge, H. et al., (1998) Nucl. Acids Res. 26 , 3208.

Martin, S.A. et al., (1975) J. Biol. Chem. 250 , 9322.Venkatesan, S. et al., (1980) J. Biol. Chem. 255

Higman, M.A. et al., (1992) J. Biol. Chem. 267 , 16430.

Myette, J.R. and Niles, E.G. (1996) J. Biol. Chem. 271 , 11936.

Barbosa, E. and Moss, B. (1978) J. Biol. Chem. 253 , 7692.

Barbosa, E. and Moss, B. (1978) J. Biol. Chem. 253 , 7698.

Schnierle, B.S. et al., (1992) Proc. Natl. Acad. Sci. USA 89 , 2897.

 Pasquinelli, A.E. et al., (1995) RNA 1 , 957.

 

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Related products

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200