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

Cat. No.: D1518364

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

Storage

Room temperature,Store at -20°C

Shipped In

Ice chest + Ice pads

Application

PCR

Panoramica

  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

Specifiche e purezza

BioReagent,Suitable for molecular biology,sterile,5×

Stabilità e conservazione

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

Condizioni di conservazione di stoccaggio

Room temperature,Store at -20°C

Spedito in

Ice chest + Ice pads

Questo prodotto richiede spedizione a catena fredda. I servizi di terra e altri servizi economici non sono disponibili.

Grado

BioReagent, Sterili, Suitable for molecular biology

Product Line

biological_products

Domande frequenti

What does Suitable for molecular biology mean?

Suitable for molecular biology indicates that the product has been processed and tested for molecular biology and proteomics. Typical specifications include low or undetectable nuclease/protease activity, low endotoxin, controlled microbial limits and high lot-to-lot reproducibility.

What does BioReagent mean?

BioReagent indicates that the product has been processed and tested for biological and biochemical applications. Specifications may include endotoxin limits, microbial controls, sterility, low DNase/RNase/protease activity, and verified biological activity where relevant.

How should this product be stored?

Store at ?20 °C. Freezer storage is required to maintain the specified shelf life.

How is this product shipped?

This product ships in an insulated container with ice pads. Unpack on arrival and transfer it to the storage condition stated above.

What documentation is provided?

Available product documentation, including Certificates of Analysis (COA), Safety Data Sheets (SDS), and specification sheets, is shown in the product document area. Document availability and access follow the current site policy.