highQu® ALLin™ HS Isothermal DNA Amplification Kit

Opis towaru

ALLin™ HS Isothermal DNA Amplification Kit enables detection of as little as 3 target DNA molecules in a short time of 20 minutes even without the use of a thermal cycler. The Kit includes a 2X master mix with optimized high-performance buffer, dNTPs and a recombinant Hot Start Bst Polymerase having strong 5’ - 3’ strand displacement activity and efficient 5’ - 3’ polymerase activity working at 55 - 70°C. The activity of the enzyme is blocked at ambient temperature due to molecular inhibition based hot-start technology, and the polymerase is activated only at 45 - 50°C what reduces non-specific amplification, primer dimer formation and is therefore excellent for an ambient temperature reaction setup. The polymerase has neither 5’ - 3’ nor 3’ - 5’ exonuclease activity and retains only minor reverse transcription activity. The kit includes PCR water; only templates and primers have to be supplied by the user. Quantitative Fluorescent Dye is included for an optional use, for a real-time detection in FAM channel on any qPCR cycler. 

ALLin™ HS Isothermal DNA Amplification Kit is a tool of choice for such applications like LAMP, WGA, RAM with an additional advantage of ambient temperature set up and higher temperature reactions, what makes amplification of complex and GC-rich templates more efficient. 

Technical characteristics of Hot Start Bst Polymerase (large fragment):

• The polymerase is blocked at ambient temperature and activated at temperatures above 45°C.
• Strong 5’ - 3’ strand displacement activity 
• 5’ - 3’ polymerase activity 
• No 5’ - 3’ exonuclease activity
• No 3’ - 5’ exonuclease (proofreading) activity 
• Minor reverse transcriptase activity (for RNA, use ALLin™ Isothermal 1Step RNA Amplification Kit (#IRK0101) which includes RTase)
• Optimal amplification temperature is 65°C. 
• Working temperature range is 55 - 70°C. 
• Optimal reaction time is 20 minutes, if needed, the reaction can be extended to 30 - 60 minutes
• The enzyme is inactivated in 10 minutes at 80°C. 
• Quantitative Fluorescent Dye has the excitation max. at 482 nm and emission max. at 512 nm.

The use of this product in certain applications may be covered by patents. The user has to analyse all applicable Limited Use Label Licenses and may need licensing for certain cases.

Applications

• Isothermal DNA amplification at elevated temperature
• Real-time detection of DNA amplification
• LAMP - loop-mediated isothermal amplification
• WGA - whole genome amplification
• RAM - ramification & RPA - recombinase polymerase amplification

Benefits

• Hot Start Bst for an ambient temperature setup and minimized background
• Efficient 20 minutes DNA amplification at constant 55-70°C temperature
• ALLin™ format, supplied with water and a dye for fast real-time detection
• Hot Start Bst DNA Polymerase with strong strand displacement activity 
• Robust on complex templates and crude samples
• Low-copy (<3 molecules) targets detection