1copy™ COVID-19 qPCR Kit

Details for Product No. ABIN6952539
Antigen
Reactivity
SARS Coronavirus-2 (SARS-CoV-2)
3
Application
Quantitative real-time PCR (qPCR)
Options
Purpose 1copy™ COVID-19 qPCR Kit provides the fast and accurate testing solution for COVID-19, specifically targeting the E gene for beta coronavirus and the RdRp gene for COVID-19 in sputum, nasopharyngeal swab and oropharyngeal swab.
Sample Type Nasal Secretions, Oral Sample, Saliva
Specificity Our COVID-19 Real-time PCR assay is based on the WHO & KCDC reference method and it has been carried out the in sillico analysis for all registered COVID-19 sequence database.
Components
  • Master mix (2 x 1000 μL)
  • Primer/Probe mix 1(E gene) (100 μL)
  • Primer/Probe mix 2(RdRp gene) (100 μL)
  • Control 1 (E gene) (100 μL)
  • Control 2 (RdRp gene) (100 μL)
  • DEPC Water Clear (1000 μL)
Material not included
  • RNase/DNase free consumables (disposable latex or vinyl gloves)
  • Filter tips
  • 0.5 mL or 0.2 mL PCR tubes or 96-well PCR plates
  • 1.5 mL micro tubes
  • Sealing film
  • Ice or cooling/cold block
  • Microliter pipettes (1~10μL,10~100μL, 100~1000μL)
  • Mini centrifuge (0.2 mL/0.5 mL tubes, 10,000 rpm) or Benchtop centrifuge with rotor for 0.2 mL /0.5 mL reaction tubes (capable of attaining 10,000 rpm), vortexer
  • Real-time PCR instrument
  • Reagents or Kit for RNA extraction
Target Name (Antigen)
Alternative Name SARS-Coronavirus-2
Application Notes Prepare RNA specimens using RNA extraction Kit or manual method for RNA extraction.
Comment

Software setting
Recommended Real-time PCR instruments

  • Roche Light Cycler® 480
  • Rotor-Gene® Q 5plex HRM
  • Applied Biosystems® Quantstudio5
  • Applied Biosystems® 7500 Real-Time PCR system
  • CFX96™ Real-Time PCR Detection system

Threshold value for each equipment are as follows
  • Light Cycler 480 = Whether to check Ct when setting
  • Rotor-Gene Q 5plex HRM = 0.3
  • Applied Biosystems® Quantstudio5 = 70,000
    ""Abs Quant/2nd Derivative Max"" is Analysis menu
  • Applied Biosystems® 7500 = 250,000
  • CFX96™ Real-Time PCR Detection system = 1,500

Assay Time 1.5 h
Assay Procedure

RT-qPCR preparation

  1. Make mixture in PCR tube according to below table.
    • Recommended to use sample of RNA 5μL/reaction
    • Use control 5μL/reaction
    • Use negative control 5μL/reaction of the provided DEPC-DW
    Number of reactions for each primer probe mix = total number of samples + 2
    - E gene primer Probe mix1 use + Control1 + Negative Control
    - RdRp gene primer Probe mix1 use + Control1 + Negative Control

    ------------------
    Mixture Components 1 reaction (Total volume : 20 μL)

    Master mix :10 μL
    Primer Probe mix: 1μL
    RNA sample: 5 μL
    (In Control, instead of RNA sample) (5 μL)
    DEPC Water: Up to 20 μL
    ------------------
  2. Gently mix the mixture by vortexing and spin the tubes.
  3. Place the samples on the 96 well PCR plate.
  4. Cover wells the sealing film and spin the plate.
  5. PCR reaction conditions follows as below.
  6. ------------------
    RT (55 °C / 25 min) 1 Cycle
    Incubation (95 °C / 5 min) 1 Cycle
    Amplification (95 °C / 10 sec) (58 °C
    / 30 sec) 45 Cycles

    *Measure florescence at 58 °C (FAM channel)
    ------------------

Calculation of Results

Experimental results confirm the failure of the experiment through the result of control and negative control.
Positive/Negative(+/-) determination of each control, negative control, and sample result is determined by whether the curve of each result crosses the threshold line after applying the threshold value for each device. (+ if cross, otherwise -)

Threshold value for each equipment are as follows

  • Light Cycler 480 = Whether to check Ct when setting
  • Rotor-Gene Q 5plex HRM = 0.3
  • Applied Biosystems® Quantstudio5 = 70,000
    ""Abs Quant/2nd Derivative Max"" is Analysis menu
  • Applied Biosystems® 7500 = 250,000
  • CFX96™ Real-Time PCR Detection system = 1,500
The interpretation of the FAM Detection Chanel results follows the table below:

E gene assay
Control: +
Negative Control: -
Sample: +
beta coronavirus detected

Control: +
Negative Control: -
Sample: -
beta coronavirus not detected

Control: +/-
Negative Control: +
Sample: +/-
Test failed / Retest

RdRp gene assay
Control: +
Negative Control: -
Sample: +
COVID-19 detected

Control: +
Negative Control: -
Sample: -
COVID-19 not detected

Control: +/-
Negative Control: +
Sample: +/-
Test failed / Retest

Restrictions For Research Use only
Handling Advice avoid freeze/thaw cycles
Storage -20 °C
Storage Comment Store the Kit below -20°C.
Expiration date for Kit is indicated on the packing box. Freezing and thawing is limited to 5 times.
Minimize the temperature difference of the components.Thaw necessary components just before using and promptly place back in freezer after use.
Supplier Images
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