Collibri™ Library Amplification Master Mix with Primer Mix, 250 reactions - FAQs

View additional product information for Collibri™ Library Amplification Master Mix with Primer Mix - FAQs (A38540050, A38540250)

11 product FAQs found

Are PCR products generated using the Collibri Library Amplification Master Mix blunt-ended?

Yes, PCR products generated using the Collibri Library Amplification Master Mix are blunt-ended.

Is Collibri Library Amplification Master Mix compatible with magnetic beads?

Yes, Collibri Library Amplification Master Mix is compatible with magnetic beads provided in Collibri library preparation kits and in Agencourt AmPure XP system from Beckman Coulter.

What is the fidelity of the Collibri Library Amplification Master Mix?

The Collibri Library Amplification Master Mix contains Platinum SuperFi DNA Polymerase that has strong 3' to 5' exonuclease (proofreading) activity for superior amplification accuracy, greater than 100-fold in comparison with Taq polymerase.

What are the recommended storage conditions for the Collibri Library Amplification Master Mix?

We recommend storing the Collibri Library Amplification Master Mix at -20 degrees C.

Is Collibri Library Amplification Master Mix suitable for amplification of libraries prepared from FFPE samples?

Yes, however, additional PCR cycles (1-3) are typically needed with FFPE DNA or other challenging samples. To determine the optimal PCR conditions, use the table in the user manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017582_Platinum_SuperFi_LibraryAmp_MM_w_Primer_Mix_UG.pdf) with the required number of PCR cycles per sample size as a starting point.

How many PCR cycles should I use for library amplification with the Collibri Library Amplification Master Mix?

To determine the optimal PCR conditions, use the table in the user manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017582_Platinum_SuperFi_LibraryAmp_MM_w_Primer_Mix_UG.pdf) with the required number of PCR cycles per sample size as a starting point. Add additional PCR cycles to amplify libraries prepared from FFPE DNA or other challenging samples.

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

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).

Is Collibri Library Amplification Master Mix compatible with bisulfite-treated DNA?

No. Collibri Library Amplification Master Mix contains Platinum SuperFi DNA Polymerase that does not incorporate dUTP and is not compatible with uracil-containing primers and templates. For this reason, Collibri Library Amplification Master Mix is not suitable for amplification of bisulfite-converted DNA.

Does Collibri Library Amplification Master Mix incorporate dUTP?

No. Collibri Library Amplification Master Mix contains Platinum SuperFi DNA Polymerase that does not incorporate dUTP and is not compatible with uracil-containing primers and templates. For this reason, Collibri Library Amplification Master Mix is not suitable for amplification of bisulfite-converted DNA.

Is Collibri Library Amplification Master Mix compatible with AmpliSeq technology?

No, this product is not designed for amplification of AmpliSeq libraries.

What is the difference between Platinum SuperFi PCR Master Mix and Collibri Library Amplification Master Mix?

Both master mixes include Platinum SuperFi DNA Polymerase that has strong 3' to 5' exonuclease (proofreading) activity for superior amplification accuracy. However, the master mix composition is different. Platinum SuperFi PCR Master Mix has been optimized for end-point PCR amplification of a variety of templates, including long and difficult samples. Collibri Library Amplification Master Mix is based on a proprietary reaction buffer that has been specifically optimized for efficient and uniform amplification of NGS libraries regardless of GC content.