Do I need to water wash or chlorite clean the Ion S5™ and Ion™ S5™ XL Systems?

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Answer

Unlike the Ion PGM™ and Ion Proton™ Systems, the sequencing reagents for the Ion S5™ and Ion™ S5™ XL Systems come with a cleaning solution bottle, which the instrument uses to automatically clean itself after each run.

Answer Id: E11456

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How many runs per initialization can I perform on the Ion S5™ and Ion™ S5™ XL Systems?

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Answer

You can perform one run per initialization.

Answer Id: E11457

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If you need to stop in the middle of the SilverQuest™ or SilverXpress™ silver staining procedure, at what point should this be done?

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Answer

The sensitizing step is the optimal point to stop the procedure. The gel can be left overnight in the Sensitizing Solution. Although some other kits recommend leaving gels in the fix step, we have found that overnight fixation diminishes stain performances.

Answer Id: E3844

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What is a suitable generic antibody starting concentration for Western blotting (WB), ELISA, immunohistochemistry (IHC), immunofluorescence (IF), immunoprecipitation (IP), or flow cytometry (FC)?

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Answer

It is recommended that you first check the product insert for any recommended usage guidelines.

For general purposes, starting concentrations are as follows:
ELISA--0.5-1.0 µg/mL
WB--1-5 µg/mL
IP--2-5 µg per reaction (1 X 10e6 cells)
IHC/IF--1-10 µg/mL
FC--10-20 µg/mL

Please note that these are general guidelines only.

Answer Id: E4942

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What is the concentration of Vial 10, ELOSA positive control?

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Answer

0.25 ng/μL

Answer Id: E14064

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I have some Ion PI™ Chips v2. Is it okay if I use them with the Ion PI™ Hi-Q™ Chef Kit chemistry?

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Answer

No. The Ion PI™ Chips v2 are not optimized for use with the Ion PI™ Hi-Q™ Chef Kit chemistry. We recommend using the Ion PI™ Chips v3, which have been pretreated during manufacturing and optimized for use with the Ion PI™ Hi-Q™ Chef Kit chemistry.

Answer Id: E11458

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Can I directly clone, propagate and express in BL21 without using TOP10?

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Answer

It is imperative that a cloning strain such as TOP10 be used for characterization of the plasmid, propagation, and maintenance. BL21 cells are wild-type for endA and recA, which could result in poor miniprep quality and a greater chance of plasmid rearrangements due to recombination. In addition, BL21 cells contain the T7 RNA polymerase gene which is expressed at low levels even in the absence of inducer. If the gene is toxic to E. coli, plasmid instability and/or cell death can result.

Answer Id: E3845

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What promoter is used to drive expression in the Invitrogen™ GeneArt™ CRISPR vectors?

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Answer

gRNA expression is driven by the Pol III U6 promoter.

Answer Id: E10152

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What is the concentration of the PHA-M solution?

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Answer

The PHA is a crude extract of red kidney beans. No protein concentration assay is performed. We recommend using the product at 1-2% by volume, or you may choose to optimize the dose by titration.

Answer Id: E4943

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I have labeled my samples with FlashTag™ labeling kit, but I am not ready to hybridize the miRNA Arrays. What should I do?

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Answer

Store the biotin-labeled RNA samples on ice for up to 6 hours, or at -20 degrees C for up to two weeks.

Answer Id: E14065

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Can I transfect adherent cells on a plate using the Neon™ transfection system?

Product FAQ

Answer

You cannot. You need to lift the cells off of the plate through a normal method such as that used to “passage” the cells (e.g., trypsin), perform the transfection in the Neon tip, and re-plate the cells in the plate after transfection.

Answer Id: E5534

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When should the ExpiCHO Feed and ExpiFectamine CHO Enhancer be added to the cultures?

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Answer

ExpiCHO Feed and ExpiFectamine CHO Enhancer should be added 18-22 hours posttransfection for best results. These solutions may be added to the flasks without prewarming. ExpiCHO Feed and ExpiFectamine CHO Enhancer may also be premixed ahead of addition to flasks to reduce the number of steps required. If using the standard protocol at 37 degrees C, it is recommended to add the feed and enhancer closer to the 18-hour timepoint.

Answer Id: E14743

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I have a second-generation lentiviral vector. Can I use it with your packaging mix?

Product FAQ

Answer

Your second-generation lentiviral vector will not be compatible with our packaging mix because our packaging mix belongs to the third generation, meaning that it does not express the tat gene; whereas your lentiviral vector will need the tat gene for virus production.

Answer Id: E9312

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What is the Master Cell Bank and Production Lot?

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Answer

When a cell line is to be used for many manufacturing cycles, a two-tiered cell banking system consisting of a master cell bank (MCB) and a working cell bank (WCB) is used.

A cell line is established from a single clone and this cell line is used to make the MCB. This MCB is characterized and extensively tested for contaminants such as bacteria, fungi, mycoplasma and viruses. Cells from the MCB are expanded to form the WCB. Each lot of cells is tested for cell growth and viability post-recovery from cryopreservation.

The establishment of cell banks for newly developed cell lines is critical to the successful development of many biological products. The cell bank system assures that the cell line is preserved, its integrity is maintained, and a sufficient supply is readily available. A well-characterized cell bank system for production of biologicals will allow for a consistent source of production cells throughout the life of the product. This method is used in many biotechnology products, which involve the use of cell lines of animal or human origin, and are at risk of viral and microbial contamination.

Here is an example of typical progression from R&D cultures to Production: R&D provides several R&D Master vials to Manufacturing. A Production Master Bank is generated (via 3-4 passages) and qualified via viability and mycoplasma testing. Most cell lines also are qualified via functional expression assay. The initial Production Lot is propagated from one vial of the Master Bank via 3-4 passages (normally taking 1-2 weeks total production time). The Production Lot is qualified via viability and sometimes, functionality. This results in a cell product that is less than 10 passages away from the original R&D clone. In most cases, no viral or mycoplasma tests are done to test the final production lot. ALL Master Cell Banks and Production Lots are stored in liquid nitrogen, therefore maximizing viability and minimizing variability between Master and Production Lots.

Answer Id: E4231

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What is the shelf life of the Ion PI™ Chips v3?

Product FAQ

Answer

The Ion PI™ Chips v3 come pretreated with a chip wash solution, and therefore have a shelf life of 9 months from manufacturing. However, please be aware that when you order the chips, time may have passed since manufacturing. To request the chips with the longest expiration date available, please contact Customer Care by phone to see which lots are in stock. Using chips that are past their expiration date can lead to a decrease in run quality.

Answer Id: E11459

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