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How long is the control PCR fragment supplied in the InsTAclone PCR Cloning kits (Cat. Nos. K1213, K1214)? Product FAQ

Answer

The control PCR fragment is 953 bp long.

Answer Id: E13234

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What is the difference between the TaqMan Universal PCR Master Mix Cat. No. 4304437 and Cat. No. 4324018? Product FAQ

Answer

Both part numbers for TaqMan Universal PCR Master Mix are supplied at a 2X concentration and contain the following components: AmpliTaq Gold DNA polymerase, dNTPs with dUTP, Passive Reference (ROX), and optimized buffer components. However, the part number 4304437 also contains AmpErase UNG and is not suitable to be used for One Step RT PCR.

Answer Id: E1581

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pCR8/GW/TOPO TA Cloning Kits QRC (Supply Centers) Manual / Product Insert

  • Version: 20 April 2012
Catalog #

Can I connect several QuantStudio 1 instruments to the same computer? Product FAQ

Answer

Yes, it is possible. Please use local area network (LAN) connection and see page 17 of the QuantStudio 1 Real-Time PCR System User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017853_QS1%20UG.pdf) for supported configuration options. However, it is not possible to connect more than one instrument directly to one computer (supplied by Thermo Fisher Scientific) using wired connection due to single Ethernet port availability on the computer supplied by us. Also, please note that starting a run simultaneously from the same computer on two connected instruments is not yet fully validated.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

Answer Id: E18831

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What volume will I receive with the Custom and Custom Plus TaqMan RNA assays for the different scales? Product FAQ

Answer

The Custom and Custom Plus TaqMan RNA assays are supplied with the following volumes:
Small: 360 µL
Medium: 750 µL
Large: 976 µL

Please note that the large scale is supplied in a 60X format, while the small and medium scales are supplied in 20X format.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

Answer Id: E16333

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Can I purchase just the 10X PCR buffer that comes with Platinum Taq? Product FAQ

Answer

The 10X PCR buffer for Platinum Taq is not available as a stand-alone item. It is only supplied as part of the enzyme kit.

Answer Id: E7625

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Where can I find the software and newest firmware for the QuantStudio 1 instrument? Product FAQ

Answer

In an effort to reduce environmental impact, we no longer supply the software on the CD. Please download it from here (https://www.thermofisher.com/uk/en/home/global/forms/life-science/quantstudio-software-download.html).

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

Answer Id: E18839

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What concentration is the vector pTZ57R/T supplied at in the InsTAclone PCR Cloning kits? Product FAQ

Answer

The vector pTZ57R/T is supplied at 55 ng/µL.

Answer Id: E13233

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How does TA Cloning work? Product FAQ

Answer

Taq polymerase has a non-template-dependent terminal transferase activity that adds a single deoxyadenosine (A) to the 3´ ends of PCR products. The linearized vector supplied in our TA Cloning kits have single, overhanging 3´ deoxythymidine (T) residues. This allows PCR inserts to ligate efficiently with the vector.

Answer Id: E4061

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With the GeneArt High-Order Genetic Assembly System, I'm getting no positive colonies detected by yeast colony PCR. Can you please offer some troubleshooting tips? Product FAQ

Answer

We would recommend trying to re-streak the colony on a fresh plate and repeat colony PCR. Do not break open the yeast cells with the beads supplied with the kit; the beads are for transformation into E. coli. Additionally, use less than 0.5 µL of diluted yeast lysate in a 50 µL PCR reaction.

Answer Id: E6862

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How are IonCode Barcode Adapters, 1153-1536 supplied? Product FAQ

Answer

IonCode Barcode Adapters, 1153-1536 are supplied in 4 separate 96-well plates containing 25 µL of each barcode:
- IonCode Barcode Adapters 1301-1396 in 96-well PCR plate (red)
- IonCode Barcode Adapters 1401-1496 in 96-well PCR plate (yellow)
- IonCode Barcode Adapters 1501-1596 in 96-well PCR plate (green)
- IonCode Barcode Adapters 1601-1696 in 96-well PCR plate (blue)

Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.

Answer Id: E18653

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What is the difference between DNase I and Amplification Grade DNase I? Product FAQ

Answer

The Amplification Grade DNase I (Cat. No. 18068-015) is subjected to an extra final HPLC purification step to remove traces of RNases. The Amplification Grade DNase I is supplied as 1 unit/µL and comes with 10X buffer (200 mM Tris-HCL pH 8.4, 20 mM MgCl2, 500 mM KCl) and a vial of 25mM EDTA.

In RT-PCR, a large excess of Amplification Grade DNase I could be used to digest an RNA template without degradation of the RNA (in-house data). Use Amplification Grade DNase I to remove genomic DNA carryover in RNA samples prior to RT-PCR.

The regular DNase I is supplied at 5-15 mg/mL (50-375 U/µL) and does not come with its own buffer.

Answer Id: E2946

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Should I use the gel extraction or the PCR purification method before cloning inserts into the pCR-XL-2-TOPO vector? Product FAQ

Answer

Large inserts should be gel-purified prior to cloning. Gel purification of long PCR products often improves efficiency, as smaller non-specific bands that may not be visible on the gel will interfere with cloning of the insert of interest. However, the TOPO XL-2 Complete PCR Cloning kit is supplied with the PureLink Quick Gel Extraction and PCR Purification Combo Kit for added flexibility allowing for faster PCR purification when inserts are smaller, while primers and dNTPs still need to be removed.

Keep in mind that the TOPO reaction does have a strong size bias, and that any smaller fragments present in the PCR reaction would clone much more efficiently than the fragment of interest, even if the smaller fragment is not readily visible on a gel. Therefore, it is highly recommended that the PCR product of interest be gel-purified.

Answer Id: E16001

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I am using the TOPO XL-2 Complete PCR Cloning Kit. Should I use the gel extraction or the PCR purification method for purification of my insert? Product FAQ

Answer

Large inserts should be gel-purified prior to cloning. Gel purification of long PCR products often improves efficiency, as smaller non-specific bands that may not be visible on the gel will interfere with cloning of the insert of interest. The TOPO XL-2 Complete PCR Cloning Kit is supplied with the PureLink Quick Gel Extraction and PCR Purification Combo Kit for added flexibility allowing for faster PCR purification when inserts are smaller, while primers and dNTPs still need to be removed.

Keep in mind that the TOPO reaction does have a strong size bias, and that any smaller fragments present in the PCR reaction would clone much more efficiently than the fragment of interest, even if the smaller fragment is not readily visible on a gel. Therefore, it is highly recommended that the PCR product of interest be gel-purified.

Answer Id: E14704

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How should I determine the optimal annealing temperature to use with the Collibri Library Amplification Master Mix? Product FAQ

Answer

An annealing temperature of 60 degrees C is recommended for the Primer Mix supplied with the kit (Cat. No. A38540050, A38540250). Optimization of annealing temperature may be required for different user-supplied adaptors and primers. Note that the annealing temperature for Platinum SuperFi DNA Polymerase is typically higher than that for other DNA polymerases. To determine the optimal annealing conditions, use the Tm calculator on our website at thermofisher.com/tmcalculator as a starting point. If necessary, use a temperature gradient to optimize further and empirically determine the ideal annealing temperature. The annealing gradient should extend up to the extension temperature (two-step PCR).

Answer Id: E17652

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