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  6. Invitrogen™ Collibri™ Library Amplification Master Mix with Primer Mix

Invitrogen™ Collibri™ Library Amplification Master Mix with Primer Mix

Catalog No. A38540250
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Catalog No. A38540250 Supplier Invitrogen™ Supplier No. A38540250
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Description

Invitrogen™ Platinum SuperFi Library Amplification Master Mix is a ready-to-use solution designed for amplification of next-generation sequencing (NGS) libraries.

Master mix includes Platinum SuperFi™ DNA Polymerase in combination with a proprietary reaction buffer containing all necessary components for efficient library amplification with minimal bias and superior accuracy.

 Features include:

  • High fidelity—minimal error rates similar to PCR-free methods
  • High efficiency—high yields from minute input amounts
  • Minimal bias—uniform amplification regardless of GC content

 

  • Supplied with a 10X Primer Mix that targets the P5 and P7 regions of Illumina adapters. The master mix is supplemented with an inert blue dye, while the primer mix contains a yellow dye. Mixing both components in a PCR reaction turns the final PCR solution green. This provides a visual aid when pipetting and helps decrease the risk of pipetting errors during reaction setup
  • Platinum SuperFi™ DNA Polymerase has both 5’ to 3’ polymerase and 3’ to 5’ exonuclease (proofreading) activities, but lacks 5’ to 3’ exonuclease activity. Exceptionally strong proofreading activity ensures amplification of NGS libraries with supreme sequence accuracy, producing NGS error rates similar to PCR-free methods
  • Proprietary reaction buffer contains all necessary PCR components and additives to enable efficient and uniform library amplification regardless of GC content, helping improve coverage across GC- and AT-rich sequences and other complex regions. High amplification efficiency enables the use of fewer cycles to achieve the desired yield
  • Platinum hot-start technology inhibits DNA polymerase activity at ambient temperatures, allowing room temperature reaction setup and storage of pre-assembled PCR reactions for up to 24 hours prior to the PCR.

Specifications

Content And Storage • 5 x 1.25 mL Collibri Library Amplification Master Mix (2X)
• 5 x 500 µL Primer Mix

Store at -20°C.
Sample Type DNA, RNA
Fidelity (vs. Taq) ≥100
GC-Rich PCR Performance High
For Use With (Application) Sequencing
Hot Start Built-In Hot Start
No. of Reactions 250
Overhang Blunt
Product Line Collibri
Product Type Library Amplification Master Mix
Quantity 250 reactions
Reaction Format SuperMix or Master Mix
Shipping Condition Dry Ice
Test Time Standard
Form Liquid
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Frequently Asked Questions (FAQs)

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.

For Research Use Only. Not for use in diagnostic procedures.