In-vitro diagnostic kit for the extraction of bacterial DNA

  • Simple 2-pipette step protocol reduces consumable costs and incubation times

  • From sample to detection in under 3 minutes

  • Isolation of DNA from Gram positive and Gram negative bacteria without Proteinase K

  • Compatible with agar, liquid broth and standard laboratory buffers such as PBS

  • Facilitates automation

Arcis Pathogen Kit

UFL004 50 Preps POA
UFL010 500 Preps POA

Need

  • Compatibility with multiple growth media
  • Protocols that can be used with multiple types of pathogens
  • Simplification that minimises need for centrifugation, heat, and enzymatic treatment
  • Safe non-hazardous chemistry

Solution

  • Standard protocol proven to work on a variety of growth media and sample types
  • Simplified workflow completely avoids centrifugation, heat, and enzymatic treatment
  • Allows rapid detection in resource limited settings
  • REACH-compatible chemistry removes safety risks

Use Arcis Sample Prep Kit to enable nucleic acid extraction from whole blood, tissue, saliva, bacteria and plant cells. Sample Prep Kit is compatible for use directly in molecular biology techniques including PCR, Next Generation Sequencing and Isothermal Amplification.

Standard Protocol

  1. If samples are frozen ensure that they have thawed completely before starting this procedure. Add 90μl of sample to 150μl of Reagent 1 (or scale up for larger sample volume). Mix thoroughly using a pipette or by vortex mixing.
  2. Incubate for one minute at room temperature. At this point DNA is stabilised for 90 days and RNA is stabilised for up to 7 days at room temperature, provided there is no further processing.
  3. Take 5μl of the above lysed mixture and combine with 20μl of Reagent 2 (or scale up for larger sample volumes maintaining the 1:4 ratio). Once processed with Reagent 2, samples should be used within 4 hours or frozen at -20 ̊C.
  4. Add appropriate volume into PCR master mix (e.g. 5μl per 25μl reaction) or continue directly to other downstream technique.

Dilute Sample Protocol

  1. When handling very dilute samples such as saliva the ratio of sample to Arcis Reagent 1 can be increased to 1:1 to avoid further dilution (90μl of sample to 90μl of Reagent 1).
  2. Samples that have been processed as in step 1 of the standard protocol can be added to Reagent 2 at 1:3, 1:2 or 1:1 ratio to reduce sample dilution. See Table.
Extract from lysis reaction (μl) Reagent 2 (μl) Ratio
5 15 1:3
10 20 1:2
20 20 1:1

Arcis Sample Prep Technology

Arcis sample prep technology is a fast and convenient 2-step protocol for preparation of nucleic acids from a variety of sample types. The nucleic acids are prepared and stabilised in 3 minutes without the need for instrumentation or complicated heating or cooling steps. The DNA and RNA extracted is suitable for direct use in PCR, qPCR and RT-qPCR reactions using standard and fast cycling protocols.

  • Simple 2-step process
  • No instrumentation required
  • Nucleic acids stabilised after 1 minute – DNA shown to be stable in Reagent 1 for up to 90 days
  • Shipped and stored at ambient temperature

Step 1

Reagent 1:

Opens up cells and releases nucleic acids in less than 1 minute. At this point the nucleic acids are released and protected by the Arcis solution, but are not in a usable state. DNA in Reagent 1 is stable for up to 90 days at room temperature.

Step 2

Reagent 2:

Binds inhibitors and enhances PCR while also relaxing the DNA into a usable confirmation.  At this point the sample should be used immediately, going straight into PCR, qPCR, RT-qPCR or sequencing.

Relaxation of DNA:

The Arcis Sample Prep system consists of 2-steps. In the first step the sample is combined with a chemical which results in powerful and controlled lysis of cells and tissue. The first step also simultaneously chelates the nucleic acid and stabilises DNA for up to 90 days at room temperature. The critical second step contains a proprietary buffer which removes the nucleic acid chelation and relaxes the DNA/RNA in preparation for PCR or RT-qPCR. The proprietary buffer has also been formulated to mop up any PCR inhibitors that may have been carried over from the first step.

Validation Data:

The Arcis Sample Prep kits were compared to market leading kits and evaluated for speed and yield. The Arcis DNA Sample Prep Kits required no instrumentation and took less than three minutes from start to finish. In contrast all the other kits required centrifugation or heating and frequently required a separate lysis step before the protocol. The processing times of the kits ranged from 20 minutes to 4 hours. The total DNA extracted from each kit was assessed using the Qubit System. The P kit yielded the most DNA, but was labour intensive and time consuming kit. The Arcis kit was comparable to the Q kit, however DNA was obtained in 3 minutes compared to 60 minutes for the Q system.

The Arcis Pathogen Kit is a ready to use kit comprising two pre-filled reagent tubes enabling extraction of nucleic acids from pathogens prior to a flexible range of downstream assays. The product is intended to be used by trained users proficient in molecular biological techniques.

Examples of applications:

Product code

UFL004, UFL010

Sample type

Bacteria grown on common microbial growth substrates

Sample input volume

Varies

Reaction time

Varies

No. of reactions

50, 500

Storage

Store at room temperature, do not freeze

Shipping conditions

Room temperature