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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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I'm getting differently sized colonies after TOP10 E. coli transformation when using the miRNA lentiviral expression system. Which one should I use? Product FAQ

Answer

Some transformants may contain plasmids in which unwanted recombination has occurred between the 5' and 3' LTR. We recommend using our One Shot Stbl3 Chemically Competent E. coli cells, as they help in stabilizing lentiviral DNA containing direct repeats, and generally give rise to fewer unwanted recombinants. We recommend screening both colony sizes; however, in general, for lentiviral plasmids the small colonies tend to be the correct clones. Large colonies may have undergone a recombination event to delete part of the plasmid, thus allowing the cells to grow faster.

Answer Id: E10026

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I'm seeing cytotoxic effects after transfection of my shRNA/miRNA construct. What is causing this? Product FAQ

Answer

You can try to scale back the amount of transfection reagent used, or use a different reagent for the transfection. Additionally, ensure that the plasmid used is pure and properly prepared for transfection.

Answer Id: E10020

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Can I freeze/thaw siRNA library stock plates? Product FAQ

Answer

Library stock plates are usually resuspended at less than or equal to 1 µM and stored at -20 degrees C or lower. Optimal storage is achieved at 10 µM stock concentrations. For Silencer siRNA, we recommend that the number of freeze/thaw cycles of the stock plates is limited to less than 10 for best results. For Silencer Select siRNA, we recommend that the number of freeze/thaw cycles of the stock plates is limited to less than 50 for best results.

Answer Id: E10038

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Are the plates barcoded? Product FAQ

Answer

Yes, each plate of the library is provided with a unique barcode. No two plates have the same barcode identifier, and we can trace back samples on any plate given that barcode. The barcode identifiers are always listed in the electronic data file shipped with each library.
The barcode is located on the short end of the plate by column 12 (centered). This label includes the barcode, a human readable definition of that barcode and a plate name that helps users order plates within a library.

Answer Id: E10056

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How should I have the siRNAs in my Invitrogen Custom siRNA Library plated? Product FAQ

Answer

For custom siRNA libraries, we can provide a number of plating options. Our most requested format is to have the last 1 or 2 columns empty of each 96-well plate, and to have different siRNAs to the same targets plated in separate plates in the same well location. The empty columns facilitate easy inclusion of any desired control in the final transfection plates. When different siRNAs to the same targets are plated in separate plates in the same well location, the siRNAs can easily be pooled with robotics or a multichannel pipettor. In addition, this type of format facilitates analysis by qRT-PCR.

Answer Id: E10067

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What is the maximum sample volume that can be used when detecting Mycoplasma using the PrepSEQ Mycoplasma Nucleic Acid Extraction kit? Product FAQ

Answer

The maximum sample volume is 50 mL (or up to 2 x 10e8 cells).

Answer Id: E10073

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What downstream applications can the purified PCR product from PureLink Pro 96 PCR Purificaiton kit be used for? Product FAQ

Answer

The purified PCR product is suitable for automated fluorescent DNA sequencing, restriction enzyme digestion, cloning.

Answer Id: E10079

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What is the sensitivity of the MicroSeq 500 kits? Product FAQ

Answer

As long as you rstart from a visible colony or cell pellet, MicroSeq kits will work.

Answer Id: E10111

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What cell lines have been tested with your mRNA nuclease CRISPR system? Product FAQ

Answer

We have tried our mRNA nuclease CRISPR system in multiple cell lines including: 293, HeLa, U2OS, HCT116, mouse neuro-2a, mouse ES cells, iPS cells, K562, Jurkat cells, CHO cells, and A549.

Answer Id: E10169

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How does Thermo Fisher Scientific treat confidential information like my gene/siRNA lists? Product FAQ

Answer

Your gene list and custom siRNA information is kept strictly confidential and is not used for any other purpose other than to prepare your price quote or to manufacture your custom siRNA library.

Answer Id: E10066

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How does the SYBR GreenER Dye Chemistry work? Product FAQ

Answer

The SYBR GreenER dye binds to double stranded DNA formed during PCR. During PCR, the polymerase amplifies the target sequence which creates the amplicon. As PCR continues, the dye binds to each new copy of double-stranded DNA, leading to more amplicons being created. The result is an increase in fluorescence intensity proportional to the amount of double-stranded PCR product produced.

Answer Id: E10078

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Is there a DNA extraction kit that you recommend using along with the MicroSeq 500 kit? Product FAQ

Answer

Yes, DNA extraction is greatly simplified by using the Applied Biosystems PrepMan Ultra Sample Preparation Reagent (Cat. No. 4322537), which can be used for all types of bacteria.

Answer Id: E10101

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What does CRISPR and CRISPR-Cas stand for? Product FAQ

Answer

CRISPR stands for clustered regularly interspaced short palindromic repeat; CRISPR-Cas (CRISPR-associated) systems are used for genome editing in various host organisms.

Answer Id: E10116

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What is the difference between NHEJ- and HDR-mediated repair? Product FAQ

Answer

Both HDR (homology directed repair) and NHEJ (non-homologous end joining) are cellular mechanisms through which double-stranded DNA lesions are repaired. When a repair template is not present, NHEJ occurs to ligate double-stranded breaks, leaving behind insertion/deletion (indel) mutations. HDR is an alternative repair pathway in which a repair template is used to copy the sequence to the double-stranded break. You can, therefore, introduce specific nucleotide changes or DNA fragments into your target gene by using HDR with a repair template.

Answer Id: E10122

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