MISSION® shRNA Custom Capabilities

Sigma-Aldrich is the exclusive distributor of the TRC shRNA library clones in DNA or lentiviral format and many custom services are available. However, our experience extends beyond the TRC collection because we contract manufacture numerous private collections and alternate libraries from high-throughput projects to large volumes. We recognize that the lentiviral RNAi technology is used in diverse applications and often with specialized robotics. A number of customizable options (including high-throughput projects, custom cloning and large-scale/high titer lentivirus) are already available to you and we will continue to consider your requests to bring you more flexibility.

Sigma provides innovative solutions for academic research, biotech research and pharmaceutical drug discovery. Our excellent systems, equipment and personnel provide the highest quality tools you require.

Customizable Options

• Custom Cloning
• Large-Scale and High Titer Lentiviral Particles
• Send Your Clones for Lentiviral Manufacturing
• High-throughput DNA and Lentiviral Manufacturing
• Custom Gene Family Sets
• QC Assays and Specifications
• Packaging Materials
• Other Options

Custom Cloning
The standard pLKO.1-puro vector for the MISSION TRC shRNA collections has broad applications and has been shown to be extremely effective in numerous publications. However, there are applications that require special purpose vectors and Sigma is working to deliver those to you through our custom cloning service. Two vectors are currently available for custom cloning (pLKO.1-puro and pLKO.1-CMV-tGFP). The new pLKO.1-CMV-tGFP vector allows for shRNA expression from a construct containing a CMV-driven TurboGFP fluorescent reporter (rather than hPGK driven puroR). We will clone shRNA sequences defined by you into our pLKO.1-puro and pLKO.1-CMV-tGFP vectors. Alternatively, we will clone any TRC shRNA sequence into the pLKO.1-CMV-tGFP vector upon request. The service includes cloning, sequencing, DNA preparation (500 ng) and lentiviral preparation (200 µl at 1 x 106 TU/ml, minimum titer via p24 assay). The DNA and viral preparations are available to be shipped 4–6 weeks from the time the order is placed. To review pricing or to submit a request for a quotation on your specific project, please use our MISSION Custom Request Submission Form.

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Large-Scale and High Titer Lentiviral Particles
Sigma has a versatile manufacturing platform for high-throughput or large-scale viral production of most HIV-1-based lentiviral vectors. The standard format of the MISSION TRC shRNA collection is 200 µl of 1 x 106 TU/ml (via p24 assay) minimum titer*. Any clone, target set, or control can be produced at larger volumes with the same minimum titer specification or higher titers. We also accommodate large-scale and high titer viral preparation using clones you send to us (must be compatible with our system). Sigma’s robust and optimized processes will save you time and reduce variability in your transduction experiments. Please see the table below for examples of our standard scale-up options. To review pricing or to submit a request for a quotation on your specific project, please use our MISSION Custom Request Submission Form.

Titer	Volume	Packaging	Min. # Particles
1 x 106 TU/ml*	1 ml	5 x 200 µl	1 x 106
1 x 106 TU/ml*	5 ml	25 x 200 µl	5 x 106
1 x 108 TU/ml	0.2 ml	4 x 50 µl	20 x 106
1 x 108 TU/ml	1 ml	10 x 100 µl	100 x 106
1 x 108 TU/ml	2 ml	20 x 100 µl	200 x 106
1 x 109 TU/ml	0.2 ml	4 x 50 µl	200 x 106
1 x 109 TU/ml	1 ml	10 x 100 µl	1000 x 106
Other scales and packaging configurations are accommodated.

*1.0 x 106 TU/ml via p24 assay is guaranteed. Our average titer is >1.0 x 107 TU/ml via p24 assay.

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Send Your Clones for Lentiviral Manufacturing
Our versatile manufacturing platform for high-throughput or large-scale viral production can be used for most HIV-1-based lentiviral vectors. Investigators are not limited to clones that we currently supply. Alternatively, clones developed in private laboratories can be sent to Sigma for viral production within our quality system. The size of the target sequence, gene sequences, as well as promoters can affect lentiviral titers during production. We will work to optimize viral production if necessary to exceed our minimum titer specifications. Submit a request for a quotation on your specific project by using our MISSION Custom Request Submission Form.

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High-throughput DNA and Lentiviral Manufacturing
Sigma-Aldrich has in-house developed technologies that generate the highest quality DNA for high-throughput DNA manufacturing. The quality and yields of our DNA are ideal for processing of shRNA clone libraries and result in the highest quality lentiviral particles available. Our titers of lentiviral particles generated in a high-throughput fashion are routinely in the range of 1 to 4 x 107 TU/ml (via p24 assay), well beyond our minimum titer specification. Requests for additional information may be routed through your local biotech sales representative or by emailing RNAi@sial.com.

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Custom Gene Family Sets
A large number of gene families have already been assembled and are available from Sigma in bacterial glycerol stock and lentiviral formats. For more information on these sets, please visit the Gene Family Sets page. Please inquire if you are interested in DNA formats or if you would like to have us assemble a specialized gene set that is not currently available. Requests may be routed through your local biotech sales representative or by emailing RNAi@sial.com.

QC Assays and Specifications
We will work with you to modify the number and types of tests performed. Please inquire with your project details. Email us at RNAi@sial.com

Packaging Materials
Options such as plate style can be modified. Please inquire with your project details. Email us at RNAi@sial.com

Other Options
Customizable options are not limited to those specified. Please inquire with your project details. Email us at RNAi@sial.com

Visit the MISSION RNAi homepage for links to a wide array of product information,
RNAi and lentiviral resources.