LRHSNTKB

KAPA LongRange

HotStart PCR Kit

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

Properties

• Quality Level: 100
• usage: sufficient for 150 reactions; sufficient for 400 reactions; sufficient for 800 reactions
• shelf life: ≤18 mo.
• feature: High Fidelity PCR; Long & Accurate PCR; dNTPs included; hotstart
• packaging: pkg of 100 U (KK3501); pkg of 250 U (KK3502); pkg of 500 U (KK3503)
• manufacturer/tradename: Roche
• technique(s): PCR: suitable
• input: purified DNA
• storage temp.: −20°C

Description

General description

The KAPA 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 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 has been used in:

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

Biochem/physiol Actions

Both KAPA Taq HotStart^® DNA polymerase and a modified archaeal (Type B) DNA polymerase enzyme 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.
• KAPA LongRange Buffer (Mg-free) is designed for long-range, sensitive PCR. MgCl₂ is supplied separately for optimization.

Quality

Each batch of KAPA LongRange HotStart DNA Polymerase is confirmed to contain <2% contaminating protein (Agilent Protein 230 Assay). The KAPA LongRange HotStart PCR system 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.

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.

Other Notes

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

Legal Information

HOTSTART is a registered trademark of Molecular BioProducts, Inc.

Kit Components Only

• KAPA Long Range HotStart^® DNA Polymerase 5 U/μL or 2.5 U/μL
• 5X KAPA Long Range Buffer
• MgCl₂ 25 mM
• dNTP Mix 10 mM each dNTP

Safety Information

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