Annealing Buffer for RNA Oligos

Aladdin's Annealing Buffer for RNA Oligos (5X) is used for annealing complementary RNA oligos to form double-stranded RNA to synthesize siRNA after cell transfection. This product can ensure proper annealing by effective prevention of RNA oligos from forming hairpin structures.This product is easy-to-use. Users just need to mix RNA oligos and the Annealing Buffer for RNA Oligos (5X) in an appropriate ratio and place the reaction in a thermocycler to complete the annealing in 60 minutes. This product is free free RNase to avoid RNA degradation.This product is sufficient for 50 reactions when a reaction volume of 100µl is used.Precautions：Annealing Buffer for RNA Oligos (5X) is for RNA oligos only, not for DNA oligos.As RNA is very susceptible to degradation, extreme care must be taken to prevent RNase contamination during the course of experimentation. 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 RNA oligos to be annealed to 200µM with DEPC-treated water. Thaw the Annealing Buffer for RNA Oligos (5X) and mix well.2.Add 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.ComponentVolumeDEPC-treated Water30µlAnnealing Buffer for RNA Oligos (5X)20µlRNA oligo A (200µM)25µlRNA oligo B (200µM)25µlTotal volume100µStepTemperatureTimeDenaturation90℃1 minuteAnnealing-0.1℃/5s, to 25℃60 minutesStorage4℃∞Note 1: The gradient temperature decreasing rate can also be set at -1℃/60s. Note 2: When a thermocycler is not available, the PCR tube can be placed in a water bath with appropriated amount of 90℃ water that can achieve the temperature decreasing from 90℃ to 25℃ within 1-2 hours naturally. However, the annealing efficacy will be inferior to that obtained using a thermocycler. 4.The annealed products can be directly used for cell transfection, or stored at -20℃ for future use.Product Citations: 1.Gao JZ, Li J, DU JL, Li XL. Long non-coding RNA HOTAIR is a marker for hepatocellular carcinoma progression and tumor recurrence. Oncol Lett. 2016 Mar;11(3):1791- 1798.2.Wu XY, Zhang CX, Deng LC, Xiao J, Yuan X, Zhang B, Hou ZB, Sheng ZH, Sun L, Jiang QC, Zhao W. Overexpressed D2 Dopamine Receptor Inhibits Non-Small Cell Lung Cancer Progression through Inhibiting NF- κB Signaling Pathway. Cell Physiol Biochem. 2018;48(6):2258-2272.3.He Q, Shi X, Zhou B, Teng J, Zhang C, Liu S, Lian J, Luo B, Zhao G, Lu H, Xu Y, Lian Y, Jia Y, Zhang Y. Interleukin 8 (CXCL8)-CXC chemokine receptor 2 (CXCR2) axis contributes to MiR-4437-associated recruitment of granulocytes and natural killer cells in ischemic stroke. Mol Immunol. 2018 Sep;101:440-449.Note: For more product citations, please visit website of this product

Specification: 5X