OnePCR reaction mixture

Details for Product No. ABIN2868533, Supplier: Log in to see
Application
Polymerase Chain Reaction (PCR)
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Brand OnePCR™
Characteristics OnePCR™ is a ready-to-use PCR reaction mixture. Simply adding primers and template, the reagent will execute primer extensions and other molecular biology applications. OnePCR™ is a pre-mixed solution containing Taq DNA polymerase, PCR Buffer, dNTPs, gel loading dyes and Novel Green. It contains the fluorescence dye, which is directly detected on the blue-light transilluminator or UV epi-illuminator afterthe DNA electrophoresis. The OnePCR™ mixture is supplied at the 1.1X concentration to allow approximately 10 % of the final reaction volume to be used for the addition of the primer and template
Application Notes Optimal working dilution should be determined by the investigator.
Assay Procedure

Standard PCR with OnePCR™
1. For each 50 μLreaction,assemble the following components in a 0.2 mL PCR tube on ice just prior to use: 45 μL OnePCR™ (=1x), variable amount of forward and reverse primer (to final conc. of 0.1-0.2 μM), DNA template (final conc. 4pg-500 ng), total volume = 50 μL
2. Mix gently. If necessary, centrifuge briefly. Cap tubes and place in the thermal cycler.
Process in the thermal cycler for 25-35 cycles as follows: Initial Denaturation: 2-5 minutes at 94 °C, Denaturation: 20-40 seconds at 94 °C, Annealing: 1 min at the proper annealing temperature (30 cycles), Extension: 2 min at 72 °C, Final extension: 5 min at 72 °C
Note: Optimal conditions for amplification will vary depending on the primers and thermal cycler used. It may be necessary to optimize the system for individual primers, template and thermal cycler.
4. After the PCR reaction, RNA DNA electrophoresis will detect the PCR product.
5. Use the UV or blue-light transilluminator or UV epi-illuminator to photograph the gel.
Note: When the DNA concentration is less than 4pg, it may cause the migratory shift when performing the electrophoresis. To remedy this observation, we recommend to conduct the following steps ( please refer to the experimental procedures), or use the PCR Clean-Up & Gel Extraction Kit to remove the Novel Green prior to post-staining with the Novel Green again for restoring the DNA molecular weight in the original position.

Removel of Novel Green:
1. Immerse the PCR product containing the Novel Green into the 100 mM NaCl and add 2.5 volumes of absolute or 95 % ethanol.
2.Incubate on ice for 20 minutes.
3.Centrifuge the mixture at 4 °C for at least 10 minutes.
4. Remove the suspension of ethanol and wash the pellet with 1 mL of 70 % ethanol.
5. Dry the residual ethanol and resuspend the double-stranded DNA in the TE.

Restrictions For Research Use only
Format Liquid
Precaution of Use May be irritating to the eyes and respiratory tract. Components of the product may beabsorbed into the body through the skin. Harm may be caused to the digestive tract if swallowed.Caution: Avoid inhalation, ingestion, and skin contacts. If swallowed, rinse mouth and throat thoroughly with water. Inhalation: If inhaled, remove victim from area of exposure. Skin contact: Wash immediately and thoroughly with plenty of water. Eye contact: In case of contact, wash immediately and thoroughly with plenty of water. Consult a doctorin the event of any complaint.
Handling Advice OnePCR™ is light sensitive and should be stored and protected from light
Storage 4 °C/-20 °C
Storage Comment Store at 4°C for up to 6 months and at -20°C for up to 1 year.
Expiry Date 12 months
Supplier Images
 image for OnePCR reaction mixture (ABIN2868533) Novel Green fluorescence dye in OnePCR Reaction Mixture (ABIN2868533): Fluorescence e...
 image for OnePCR reaction mixture (ABIN2868533) OnePCR Reaction Mix (ABIN2868533); PCR Mastermix for quick and easy PCR.