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View additional product information for Collibri™ Library Amplification Master Mix with Primer Mix - FAQs (A38540050, A38540250)
11 product FAQs found
Yes, PCR products generated using the Collibri Library Amplification Master Mix are blunt-ended.
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.
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.
We recommend storing the Collibri Library Amplification Master Mix at -20 degrees C.
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.
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.
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).
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.
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.
No, this product is not designed for amplification of AmpliSeq libraries.
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.