What is the difference between the Microporator (MP-100) from Digital Bio/NanoEntek and the Neon™ Transfection System?

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The Neon™ Transfection system is the second generation of the Microporator (MP-100) from Digital Bio/NanoEntek. The unique feature of both these instruments is that they use an electronic pipette tip as a transfection chamber instead of a standard electroporation cuvette. The performance of these instruments is comparable, but there are some differences. The main difference is in the user interface. The Neon™ Transfection System unit has updated firmware and can upload more programs than the Digital Bio Microporator MP-100. The MP-100 Microporator is white in color and has a smaller screen than the Neon™ Transfection System unit. The Neon™ Transfection Pipette Station is not compatible with the MP-100 pipette (white in color). This is because the sensor connector (it is a 2-pin connection) is different from that in the Neon™ Pipette Station (12-pin connection). We do not carry an adapter to allow compatibility. The kits and components in both systems are exactly identical except that they are branded as Neon™ in the Neon™ Transfection System. The old manual can be used for the Neon™ Transfection System instrument. More information regarding the history of the Neon™ Transfection System can be found here (https://www.thermofisher.com/us/en/home/life-science/cell-culture/transfection/transfection---selection-misc/neon-transfection-system/history-of-the-neon-transfection-system.html).

Answer Id: E9078

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How does the Neon™ system compare with the Amaxa Nucleofector™ II?

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Learn more about the Neon™ Transfection System versus the Amaxa Nucleofector™ II System here (https://www.thermofisher.com/us/en/home/life-science/cell-culture/transfection/transfection---selection-misc/neon-transfection-system/neon-vs--amaxa.html).

Answer Id: E9079

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What is the advantage of your Neon™ transfection chamber design over standard cuvettes?

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Unlike standard cuvette-based electroporation chambers, the Neon™ system uses a patented biologically compatible pipette tip chamber. The design of a gold-coated wire electrode inside a pipette tip has been shown to produce a more uniform electrical field and a lower pH gradient across the cell suspension. Therefore, this design allows for better maintenance of physiological conditions, resulting in very high cell survival compared to conventional electroporation (Kim JA, Cho K, Shin MS, et al. (2008) A novel electroporation method using a capillary and wire-type electrode. Biosens Bioelectron 23(9):1353–1360).

Answer Id: E9080

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Can I use the Neon™ electroporation technology for cell fusion applications?

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Cell fusion may occur "accidentally" as a side-effect during transfection using the Neon™ system, for some cell-types that tend to form cell clusters (e.g.. PC-12 cells), but unfortunately, we do not offer a Neon™ program that is optimized for cell fusion applications.

Answer Id: E9098

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Can I use my old Invitrogen™ Quant-iT™ Kits labeled “for use with Invitrogen™ Qubit™ Fluorometer” with the Invitrogen™ Qubit™ 2.0 and 3.0 Fluorometers?

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Yes, these kits will work with all Qubit Fluorometers.

Answer Id: E10255

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How does the Neon™ Transfection System compare to other delivery methods (i.e., lipid-based transfection, viral delivery)?

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The Neon™ Transfection System efficiently delivers nucleic acids (DNA/siRNA) into all mammalian cell types. In comparison, lipid-based transfection reagents are less efficient for delivering nucleic acids into hard-to-transfect cells like primary cells, stem cells, and hematopoietic cells. Viral delivery such as those using lentivirus and adenovirus can efficiently deliver DNA and siRNA from our shRNA or miRNA vectors into difficult-to-transfect cells, but producing the virus is a time-consuming and more complex process.

Answer Id: E9081

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Which tissues and cell types have been used with the NE-PER Reagents?

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We have extracted, soluble nuclear proteins from calf liver tissue, and cultured epithelial, fibroid, kidney, liver and brain cells. Many references have cited the use of various other cells and tissues with the NE-PER Reagents.

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I am running the E-Gel™ 96 High Range DNA markers. How many bands should I resolve and what percent agarose gel should I use?

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The high range DNA marker contains 5 bands of equal intensity. This marker should be run on a 1% agarose gel. On a 2% agarose gel the top two bands will not resolve.

Answer Id: E3868

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How can I run the Windows 2000 autoscheduler to run autosynch between Vector NTI™ software and LabShare™ software?

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(1) The autosync utility must be configured first. To set it up, start autosync manually. Select the desired data source and check 'Save Session Settings on Exit' box. Exit the application. It will start synchronization automatically next time it is run.

(2) Create the new Windows Scheduler task. Browse to the Vector NTI™ 9 program files directory (usually c:\program files\informax\vector nti suite 9 and select the 'VntiLsSynch.exe' application. Select the desired run frequency, start date/time, etc. When prompted by the 'Scheduled Task Wizard' for the user name/password, enter the Windows account name/password (NOT the one for LabShare#&146; program).

(3) In order to be able to reset the settings without starting the upsynchronization process automatically, run the utility manually with /c command-line switch: Start Menu->Run->\VntiLsSynch.exe /c

Please note that since Labshare™ software is a discontinued product, we are no longer supporting it.

Answer Id: E4474

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Can I use the Neon™ Transfection System for electroporation of cells other than mammalian cells? How about bacterial cells?

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Currently, we do not offer a Neon™ protocol for electroporation of bacterial cells. However, the Neon™ Transfection System may be used for electroporation of Chlamydomonas. Please refer to page 18 in the GeneArt™ Chlamydomonas Protein Expression Kit manual (https://tools.thermofisher.com/content/sfs/manuals/geneart_chlamydomonas_protein_expression_man.pdf) or page 15 in the GeneArt™ Chlamydomonas Engineering Kit manual (http://tools.thermofisher.com/content/sfs/manuals/geneart_chlamy_kits_man.pdf) or page 20 in the GeneArt™ Chlamydomonas TOPO™ Engineering Kit manual (http://tools.thermofisher.com/content/sfs/manuals/geneart_chlamy_topo_kits_man.pdf) for electroporation.

Answer Id: E9099

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How many lines of data can the Invitrogen™ Qubit™ Fluorometers store?

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The Invitrogen™ Qubit™ 3.0 Fluorometer can store up to 1,000 samples' worth of data in a .csv file.
The Invitrogen™ Qubit™ 2.0 Fluorometer can store up to 200 lines of data in a .csv file.
The original Invitrogen™ Qubit™ (1.0) Fluorometer can store up to 20 lines of data in a .csv file.

Answer Id: E10256

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After isolation of mRNA using Dynabeads™ magnetic beads and before doing reverse transcription, should I incubate the beads with bound mRNA attached to the primer, at 65 degrees C for 5 min as suggested in my reverse transcription protocol or should I just move on to cDNA synthesis (with incubation at 50 degrees C and then 65 degrees C?

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Answer

The purpose of this step (heating at at 65 degrees C for 5 min) is to open up secondary structures in the RNA. If you want to use the Oligo(dT)25 on the beads as primers for your cDNA synthesis and generate solid-phase cDNA, you should omit this step. Start with 50 degrees C (otherwise the mRNA will fall off the beads) then proceed to the 65 degrees C step.

Answer Id: E6081

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Should any reagents be avoided when using the SuperSignal West Dura Chemiluminescent Substrate?

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Yes. Avoid using sodium azide during and after probing steps involving horseradish peroxidase (HRP), as this will inhibit HRP activity. Sulfide, cyanide, fluoride and superoxide ions also inhibit HRP to some extent.

Answer Id: E8499

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What is the difference between the Neon™ Resuspension and Electrolytic Buffers?

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The Resuspension Buffer (Buffer R or T) is used to resuspend the cells prior to electroporation, whereas the Electrolytic Buffer (Buffer E or E2) is used for electroporation and is added to the Neon™ tube prior to electroporation.

Answer Id: E9082

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Can I use the NE-PER Reagents for both adherent and suspension cells?

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Yes. The kit was optimized for a packed-cell volume; therefore, adherent cells require harvesting, either by scraping or trypsinization, from the culture flask for maximum recovery.

Answer Id: E8194

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