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Regulatory requirements vary by product. Please see product-specific literature to determine product use.

How do I order miR RNAi? Product FAQ

Answer

Please visit our BLOCK-iT RNAi Designer and select miR RNAi as your target design option. This miR RNAi can then be cloned into the pcDNA6.2-GW/miR and pcDNA6.2/EmGFP-miR vectors.

Answer Id: E10001

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With regard to the promoter, H1 and U6, for the BLOCK-iT Lentiviral RNAi Expression System, is there a difference in knockdown? Product FAQ

Answer

Constitutive knockdown is virtually identical for these two promoters in HEK 293 cells. In other cell types there are reports that either H1 or U6 may be more active, though in general, differences are minimal.

Answer Id: E10009

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After ligation and transformation of my CRISPR plasmid, how should I analyze my transformants? Product FAQ

Answer

Confirm the identity of the ds oligonucleotide insert in positive transformants by sequencing. Analyze each CRISPR nuclease construct to verify:

- That the ds oligonucleotide insert is present, and in the correct orientation
- That the ds oligonucleotide insert has the correct sequence

Note: Restriction analysis is not recommended due to the small size of the ds oligonucleotide insert.

Answer Id: E10148

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How can I tell if my CRISPR genome editing experiment is working? Product FAQ

Answer

Cleavage efficiency can be detected using the Invitrogen GeneArt Genomic Cleavage Detection Assay. This assay relies on mismatch detection endonucleases to detect insertions and deletions (indels) generated during cellular NHEJ repair.

Answer Id: E10154

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How efficient is the Cas9 mRNA format compared to the all-in-one CRISPR plasmid format? Product FAQ

Answer

In most cell types tested, this complete RNA format exhibits higher cleavage efficiency than the plasmid format. Additionally, the Cas9 mRNA format circumvents the need for cloning, has a smaller payload size, allows Cas9-to-gRNA dosage optimization, flexibility with multiplexing, and does not have any promoter constraints.

Answer Id: E10160

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How does the GeneArt Genomic Cleavage Selection Kit work? Product FAQ

Answer

The vector in the kit has an OFP gene that is interrupted by the insertion of a cloning site for the target sequence of the programmable nuclease. The upstream sequence coding for the N-terminal portion of the OFP gene contains a region complementary to the 5′ end of the C-terminal region of the OFP gene. A stop codon after the N-terminal OFP sequence ensures no expression of the reporter prior to nuclease activity. The CD4 gene is out of frame for expression when the OFP gene is interrupted by the cloning site. Double-stranded breaks cause the complementary strands of the end sequences of the OFP gene to recombine, and OFP expression is restored. The CD4 gene is now in frame for expression and can be screened for via FACs analysis or Dynabeads magnetic bead selection.

Answer Id: E10173

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What is the peak excitation and emission of OFP? What type of laser should I use? Product FAQ

Answer

OFP has peak excitation of 548 nm and emission of 560 nm. A 488 nm laser is recommended for efficient excitation. Standard 530/30, 574/26 and 603/48 emission filters are recommended for detection.

Answer Id: E10179

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My results do not match data from the GeneArt Genomic cleavage selection assay. Why is this? Product FAQ

Answer

Results are locus-dependent. We recommend using the Invitrogen GeneArtGenomic Cleavage Detection Kit (Cat. No. A24372) to verify cleavage on the endogenous genomic locus.

Answer Id: E10191

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How fast can I get TAL constructs made so that I can do my KO or KI experiments? Product FAQ

Answer

Manufacturing takes place typically within 2 weeks after your order has been received.

Answer Id: E10204

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When should I use a native DNA-binding domain compared to a truncated one for TALs? Product FAQ

Answer

Truncated TAL Fok1: Recommended for mammalian cells

Native TAL Fok1: Recommended for plants

Native Activators: Higher performance in non-mammalian systems

Truncated MCS: Removing endogenous activator activity

Answer Id: E10210

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How can I quantify cleavage efficiency using the GeneArt Genomic Cleavage Detection Kit ? Product FAQ

Answer

Samples are typically run on an agarose gel, such as an Invitrogen E-Gel EX gel, followed by analysis with image software or by microfluidic electrophoresis.

Answer Id: E10216

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How much is 10 GB of storage on the Cloud? Product FAQ

Answer

This is roughly the amount of data generated by the analysis of 140 Applied Biosystems OpenArray plates.

Answer Id: E10234

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With the Applied Biosystems QuantStudio 6 and 7 Flex Real-Time PCR System Software, v1.0, I get the error message "Unable to start the run" when starting a new run. What can I do? Product FAQ

Answer

Please uncheck and check the “Data Collection” camera icon in the Run Method, and the “Start Run” button should now be active again.

Answer Id: E10228

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Do you offer stand alone replacement buffers for your specialized RNA purification kits? Product FAQ

Answer

Here (http://www.thermofisher.com/search/global/searchAction.action?query=dna%E2%81%84rna%20purification%20replacement%20buffers&resultsPerPage=15&personaFilterTerm=Catalog&show_seproductcategorynavigator=true&show_productcategorynavigator=true&show_taxonomynavigator=true&show_brandnavigator=true&firstSearch=true&mode=and&navigator=seproductcategorynavigator&modifier=General+Buffers%2FDNA%26frasl%3BRNA+Purification+Replacement+Buffers&ICID=ta-cat-dna%E2%81%84rna%20purification%20replacement%20buffers-DNA%E2%81%84RNA%20Purification%20Replacement%20Buffers) is a list of DNA/RNA Purification replacement buffers we offer as standalone.

Answer Id: E10246

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Since you now offer the Invitrogen Qubit 4.0 Fluorometer, will the Invitrogen Qubit&trade 3.0 Fluorometer be discontinued? Product FAQ

Answer

Yes.

Answer Id: E10253

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