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KAPA LongRange

HotStart ReadyMix^™, suitable for PCR, with dye

• Synonym(s): PCR
• NACRES: NA.55
• UNSPSC Code: 41106300

Properties

• usage: sufficient for 100 reactions, sufficient for 500 reactions
• shelf life: ≤18 mo.
• feature: Long Range PCR, dNTPs included, hotstart
• packaging: pkg of 1.25 mL (100 x 25 μL rxn; KK3601), pkg of 6.25 mL (500 x 25 μL rxn; KK3602)
• manufacturer/tradename: Roche
• technique(s): PCR: suitable, whole genome amplification: suitable
• input: purified DNA
• storage temp.: −20°C

Description

General description

The KAPA HotStart^® ReadyMix^™ with dye Long Range PCR system is a blend of Taq DNA polymerase and an engineered archaeal (B-family) DNA polymerase possessing proofreading capability. This two-enzyme system is designed specifically to support long-range and/or sensitive PCR, and possess 3 – 4X higher fidelity than Taq polymerase. In the hot-start formulation, KAPA Long Range is combined with a proprietary antibody that inactivates the enzyme until the first denaturation step, eliminating spurious amplification products and increasing reaction efficiency and sensitivity.

KAPA HotStart ReadyMix with dye Long Range is also supplied in 2X ReadyMix format with loading dye that contains all the components required for PCR except primers and template. The loading dye in the reaction buffer allows you to load PCR products directly onto the agarose gel with no extra steps.

Application

KAPA LongRange HotStart^® ReadyMix^™ with dye has been used for:

• Amplification of long targets and/or low amounts
• Standard short- and mid-range polymerase chain reaction (PCR)
• T-A cloning
• Targeted sample enrichment for next-generation DNA sequencing
• Complete genome amplification

Biochem/physiol Actions

Both KAPA Taq HotStart^® DNA polymerase and a modified archaeal (Type B) DNA polymerase possess 5′→3′ polymerase activity, but only KAPA Taq HotStart DNA polymerase possesses 5′→3′ exonuclease activity and only the Type B DNA polymerase possesses 3′→5′ exonuclease (proofreading) activity. The fidelity of KAPA LongRange HotStart is 2–4 fold higher than that of Taq DNA Polymerase but is lower than the fidelity of pure proofreading polymerases such as KAPA HiFi. Reaction products generated with KAPA LongRange HotStart are polyA-tailed, and can be cloned into TA-cloning vectors.

Features and Benefits

Amplify long targets with high sensitivity:

• Amplification of targets with high sensitivity and specificity up to 15 kb
• High yields with low input DNA

Outperforms with complex targets:

• Exhibits high yield, greater sensitivity and specificity where competitor long-range systems fail

Quick Notes:

• KAPA LongRange HotStart is a blend of KAPA Taq HotStart and a proofreading DNA polymerase.
• Suitable for short-, medium- and long-range PCR, and PCR with low amounts of template DNA.
• Fidelity is 2–4 times better than Taq.
• For short amplicons, replace Taq DNA polymerase for improvements in yield and/or fidelity.
• Amplify mid- to long-range targets with high yield and sensitivity.
• 2X ReadyMix with dye contains two inert tracking dyes to allow loading of PCR products directly onto agarose gels for analysis.

Preparation Note

Always ensure that the product has been fully thawed and mixed before use. Reagents may be stored at 4°C for short-term use (up to 1 month). Return to -20°C for longterm storage.

Analysis Note

Each batch of KAPA LongRange HotStart DNA Polymerase is confirmed to contain <2% contaminating protein (Agilent Protein 230 Assay). The 2X KAPA LongRange HotStart ReadyMix is subjected to stringent quality control tests, is free of contaminating exo- and endonuclease activity, and meets strict requirements with respect to DNA contamination levels.

Other Notes

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

Legal Information

HOTSTART is a registered trademark of Molecular BioProducts, Inc.

ReadyMix is a trademark of Sigma-Aldrich Co. LLC

Kit Components Only

• KAPA Long Range DNA Polymerase or KAPA Long Range HotStart^® DNA Polymerase 5 U/μL or 2.5 U/μL
• 1X KAPA Long Range Buffer
• dNTP Mix 0.3mM of each dNTP at 1X
• 2X KAPA LongRange HotStart^® ReadyMix with dye
• MgCl₂ at 1X 2 mM

Safety Information

• Storage Class: 12 - Non Combustible Liquids
• WGK: WGK 1
• Flash Point (°F): does not flash
• Flash Point (°C): does not flash