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SHC003

MISSION® pLKO.1-puro-CMV-TurboGFP Positive Control Plasmid DNA

Green fluorescent protein marker to monitor transduction efficiency

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MISSION® Control Vectors

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Talla/SKUDisponibilidadPrecio
1 pkg
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639,00 €

Acerca de este artículo

UNSPSC Code:
41106609
NACRES:
NA.51
MDL number:
Concentration:
500 ng/μL in TE buffer; DNA (10μg of plasmid DNA)

639,00 €

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Quality Segment

product line

MISSION®

concentration

500 ng/μL in TE buffer; DNA (10μg of plasmid DNA)

shipped in

dry ice

storage temp.

−20°C

General description

The MISSION® TurboGFP Control Vector is an 8347 base pair lentivirus plasmid vector that contains a gene encoding TurboGFP, under the control of the CMV promoter. The TurboGFP Control Vector is useful as a positive transfection/transduction control in experiments using the MISSION shRNA library clones. TurboGFP is an improved variant of the green fluorescent protein copGFP cloned from copepoda Pontellina plumata.

Ampicillin and puromycin antibiotic resistance genes provide selection in bacterial or mammalian cells respectively. In addition, self-inactivating replication incompetent viral particles can be produced in packaging cells (HEK293T) by co-transfection with compatible packaging plasmids. The TurboGFP Control Vector is provided as 10 μg of plasmid DNA in Tris-EDTA (TE) buffer at a concentration of 500 ng/μl.

Application

To see more application data, protocols, vector maps visit sigma.com/shrna.
MISSION® pLKO.1-puro-CMV-TurboGFP Positive Control Plasmid DNA has been used to construct GATA2 overexpression vector (oeG2), by removing the GFP cassette of this vector at the 5′ NheI and 3′ PstI cloning sites and replacing it with the full-length GATA2 transcript sequence.

Legal Information

Use of this product is subject to one or more license agreements. For details, please see http://sigmaaldrich.com/missionlicense.
MISSION is a registered trademark of Merck KGaA, Darmstadt, Germany
TurboGFP is a trademark of Evrogen Co.

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Este artículo
SHC204SHC007SHC014
description

Green fluorescent protein marker to monitor transduction efficiency

description

shRNA sequence targeting tGFP

description

shRNA sequence targeting luciferase

description

Contains a gene encoding TurboGFP, under the control of the UbC promoter

product line

MISSION®

product line

MISSION®

product line

MISSION®

product line

MISSION®

concentration

500 ng/μL in TE buffer; DNA (10μg of plasmid DNA)

concentration

500 ng/μL in TE buffer; DNA (10μg of plasmid DNA)

concentration

500 ng/μL in TE buffer; DNA (10μg of plasmid DNA)

concentration

500 ng/μL in TE buffer; DNA (10μg of plasmid DNA)

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

shipped in

dry ice

shipped in

dry ice

shipped in

dry ice

shipped in

dry ice

Quality Level

200

Quality Level

100

Quality Level

200

Quality Level

200


Clase de almacenamiento

12 - Non Combustible Liquids

wgk

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable



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Preguntas

1–9 de 9 Preguntas  
  1. What is the appropriate concentration of ampicillin to use when propagating the plasmid with DH5α-T1R?

    1 respuesta
    1. Unfortunately, there is currently not a product offering that is a DH5α-T1R competent cell line.

      Kindly reference the documentation for your cells. The response curve is cell dependent not plasmid dependent. The plasmid only selects for which antibiotic the transfected cells will survive.

      For the shRNA glycerol stocks, we supply them at TB with carbenicillin at 100 ug/ml. Kindly review our Technical Bulletin at this link:

      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/259/692/shclngbul.pdf

      ¿Le ha resultado útil?

  2. Which concentration of Ampicilin should I use to do the selection of bacterial cells containing the MISSION® Control Vector Purified DNA TurboGFP™ plasmid?

    1 respuesta
    1. For standard practice, a concentration of 100 µg/mL ampicillin is recommended for bacterial selection. For improved plasmid stability, 100 µg/mL carbenicillin is also commonly recommended as an alternative to ampicillin.

      ¿Le ha resultado útil?

  3. Is it possible to use the MISSION® pLKO.1-puro-CMV-TurboGFP™ Positive Control Plasmid DNA as an empty vector control in the experiments which contains gene of interest in pLKO.1-puro-CMV-TurboGFP vector ?

    1 respuesta
    1. This product can serve as an empty vector control only if the target backbone expresses TurboGFP.

      ¿Le ha resultado útil?

  4. dear officer I'd like to know if I'm able to clone in shRNA of choice downstream of the U6 promoter of this plasmid (SHC003)? is there any RE site available to do so? Thank you

    1 respuesta
    1. No. The cloning site, EcoR1, is ablated during production to prevent cloning into the vector. Please see the link below to review the product datasheet:
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/396/419/shc003bul.pdf

      ¿Le ha resultado útil?

  5. How much plasmid DNA do I receive when I order MISSION® Control Vector Purified DNA TurboGFP™, Product SHC003?

    1 respuesta
    1. Each MISSION Control Vector is provided as 10 μg purified plasmid DNA at a concentration of ~500 ng/μL in 10 mM Tris-HCl, pH 8.0, containing 1 mM EDTA.

      ¿Le ha resultado útil?

  6. What competent cells can I use to propagate the MISSION® Control Vector Purified DNA TurboGFP™ plasmid, Product SHC003?

    1 respuesta
    1. We use GC5 cells (identical genotype to DH5α-T1R) to propagate the Mission shRNA plasmids and controls.

      ¿Le ha resultado útil?

  7. Can I use MISSION® Control Vector Purified DNA TurboGFP™ plasmid, Product SHC003, to produce lentiviral particles?

    1 respuesta
    1. The MISSION® Lentiviral Packaging Mix can be used if you wish to produce virus in your own lab. The product code is SHP001. This product is an optimized formulation of two plasmids expressing the key HIV packaging genes and a heterologous viral envelope gene. The Lentiviral Packaging Mix is designed to be co-transfected along with a compatible lentiviral transfer vector (such as SHC003) into HEK293T cells in order to create high-titer pseudo-typed lentiviral particles used for downstream transduction applications. The procedure is available in the Technical Bulletin for product SHP001.

      ¿Le ha resultado útil?

  8. Can I purchase lentiviral particles produced using MISSION® Control Vector Purified DNA TurboGFP™ plasmid, Product SHC003, directly from Sigma-Aldrich?

    1 respuesta
    1. Yes. The product code is SHC003V.

      ¿Le ha resultado útil?

  9. What is the Department of Transportation shipping information for this product?

    1 respuesta
    1. Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.

      ¿Le ha resultado útil?

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