Annealing Buffer for DNA Oligos (5×) - BioReagent,Suitable for molecular biology,sterile,5×

Cat. No.: D1518364

GRADE & PURITY Suitable for molecular biology, BioReagent, Sterile, 5×

Storage

Room temperature,Store at -20°C

Shipped In

Ice chest + Ice pads

Application

PCR

Overview

  Annealing Buffer for DNA Oligos is suitable for routine DNA oligo annealing, and is particularly effective for difficult-to-anneal DNA oligos used in RNAi/siRNA plasmid construction. Such DNA oligos often contain approximately 20 bp complementary sequences, making them prone to forming hairpin structures that can interfere with correct pairing and annealing of the two strands.

  Annealing Buffer for DNA Oligos (5×) effectively inhibits the formation of DNA oligo hairpin structures. The annealed products can be used directly in subsequent steps such as restriction enzyme digestion followed by plasmid ligation and transformation, significantly improving the positive clone yield. This reagent is easy to use; simply mix the oligos with the annealing buffer in the appropriate proportions and complete the annealing process via a PCR instrument programmed for controlled cooling over approximately 90 minutes.

  This product is for research use only. Not for use in diagnostic procedures or other purposes.

D1518364	Component	1 mL	Storage
D1518364A	Annealing Buffer for DNA Oligos (5×)	1 mL	-20℃.
D1518364B	Nuclease-Free Water	5 mL	RT.

Procedure (For Reference Only)

1. Reagent Preparation

• Prepare the DNA oligos to be annealed as a 50 μM working solution using sterile ultrapure water or Nuclease-Free Water.

• Thaw and thoroughly mix the Annealing Buffer for DNA Oligos (5×).

2. Annealing Reaction Setup (Total Volume 100 μL)

Prepare the reaction mixture according to the table below, adding components in the order listed and mixing well. If the PCR instrument does not have a heated lid, overlay with Mineral Oil to prevent evaporation.

Component	Volume
Nuclease-Free Water	40 μL
Annealing Buffer for DNA Oligos (5×)	20 μL
DNA oligo A (50 μM)	20 μL
DNA oligo B (50 μM)	20 μL

3. PCR Instrument Annealing Program

Step	Temperature	Time	Purpose
1	95℃	2 min	Complete denaturation of oligos
2	Decrease 1℃ every 90 s to 25℃	~90 min	Programmed annealing
3	4℃	Hold	Temporary storage

4. Subsequent Use

• The annealed product can be used directly in ligation reactions or stored at -20℃ for later use.

• If subsequent enzymatic reactions (e.g., restriction enzyme digestion) are required, it is recommended to purify the annealed product first, or ensure that the volume of the annealed product constitutes less than 5% of the final reaction system to avoid buffer interference with enzyme activity.

Notes

1. Avoid repeated freezing and thawing of the reagent to prevent decreased annealing efficiency.

2. This product is specifically designed for DNA oligo annealing and should not be used for RNA oligos.

3. Please wear laboratory appropriate attire and disposable gloves during operation to ensure safety.

4. Use the reagent as soon as possible after opening to ensure optimal performance.

Specifications

Specifications & Purity

BioReagent, Suitable for molecular biology, sterile, 5×

Stability And Storage

Each component has a shelf life of 1 year under corresponding storage conditions.

Storage

Room temperature, Store at -20°C

Shipped In

Ice chest + Ice pads

This product requires cold chain shipping. Ground and other economy services are not available.

Grade

BioReagent, Sterile, Suitable for molecular biology