Annealing Buffer for DNA Oligos

Item Number

A743817

• Specifications & Purity: 5X
• Stability And Storage: Store at -20℃ for up to 1 year.
• Storage Temp: Store at -20°C
• Shipped In: Ice chest + Ice pads

Product Description

Aladdin's Annealing Buffer for DNA Oligos (5X) can not only be used for the annealing of conventional DNA oligos, but also for DNA oligos that are difficult to anneal for RNAi plasmid construction. DNA oligos used for RNAi plasmid construction usually contains about 20 complementary sequences that are easy to form hairpin structures, thus affecting the correct annealing of DNA oligos. The use of this product can effectively prevent DNA oligos from forming hairpin structures to ensure correct DNA oligo annealing and enable high efficient ligation with digested plasmids subsequently.This product is easy to use. Users just need to mix DNA oligos with this product in an appropriate ratio, and place the mix in a thermocycler to complete the annealing in about 90 minutes.This product is sufficient for 50 annealing reactions with a reaction volume of 100μl.

Precautions：

This product is for DNA oligos only, and can not be used for annealing of RNA oligos.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.

Instructions for Use：

1. Dilute the DNA oligo to be annealed to 50µM in sterilized Milli-Q water or ddH2O. Thaw the Annealing Buffer for DNA Oligos (5X) and mix well before use. 2. Set up the reaction by adding the reagents in order as follows and mix well. If the thermocycler used does not have a heated lid, add one drop of mineral oil to prevent evaporation.Nuclease-free Water 40µl Annealing Buffer for DNA Oligos (5X) 20µlDNA Oligo A (50µM) 20µlDNA Oligo B (50µM) 20µlTotal Volume 100µl3. Transfer the reaction tube to a thermocycler and perform the annealing condition as follows:StepTemperatureTimeDescription195ºC2 minutesOligo denaturation20.1ºC /8s, to 25 ºC~ 90 minutesAnnealing34ºC∞StorageNote 1: Using Bio-Rad's T100 as an example, set up the program as follows: 1. 95ºC, 2:00; 2. 95º, 0.08, -0.1ºC per cycle; 3. GOTO step 2, 700X; 4. 4ºC, ∞.Note 2: If the thermocycler available could not reduce the temperature by 0.1ºC, it can also be set to decrease at 1ºC every 90 seconds. Using Bio-Rad's T100 as an example, set up the program as follows: 1. 95ºC, 2:00; 2. 95ºC, 1:30, -1ºC per cycle; 3. GOTO step 2, 70X; 4. 4ºC, ∞.Note 3: When a thermocycler is not available, the PCR tube can be placed in a 95ºC water bath that can naturally cool down to 25ºC within 1-2 hours. However, the annealing efficacy will be inferior to that obtained using a thermocycler.4. The annealed products can be used directly for ligation, or stored at -20ºC for future use. For other applications such as enzyme digestion, the annealed products should be purified with a purification kit, or should not exceed 5% of the reaction volume to avoid the interference of the Annealing Buffer.