UltraBioâ„¢ SYBR Green qPCR Mix (C007B-566181)

Aladdin's UltraBio™ SYBR Green qPCR Mix (2X, Low ROX, UDG) is a new type of high-quality carryover prevention supermix for real-time fluorescence quantitative PCR (qPCR). It is mainly used in gene expression studies and genomics applications, with high specificity and sensitivity. This product contains an optimized ratio of high-quality UDG enzyme and dUTP to effectively eliminate false positives or low CT values caused by carryover contamination in PCR. Uracil-DNA Glycosylase (UDG), also known as Uracil-N-glycosylase (UNG), catalyzes the hydrolysis of the N-glycosidic bond between the uracil (dU) base and the deoxyribose moiety in uracil-containing DNA strands, thereby releasing free uracil. Its main application is to eliminate carryover contamination in PCR. The working principle is as followsPrecautions：PrecautionsPrecautions should be taken to avoid cross-contamination. Discard PCR products after sealing to avoid their contamination of experimental surroundings.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.Prepare qPCR reactionsa.Thaw and mix well the SYBR Green qPCR Mix (2X, Low ROX, UDG). Keep it on ice after thawing until use.b.Assemble qPCR reactions on ice as follows (e.g., in a 96-well plate):ReagentVolume (μl) SYBR Green qPCR Mix (2X, Low ROX, UDG)10Forward and Reverse Primer Mix (3μM each)2Template DNA2RNase-free Water6Total Volume20Note 1: Primer at a final concentration of 0.2-0.5μM normally works well, but primer concentration can be optimized between 0.1-1.0μM.Note 2: Add 1-10ng of cDNA or 10-100ng of genomic DNA in 20µl of qPCR reaction as a starting point. If necessary, the DNA templates can be diluted in a gradient manner to determine the optimal amount. When the cDNA obtained by RT-PCR reaction is directly used as the template, the addition amount should not exceed 10% of the total volume of PCR reaction.Note 3: A reaction volume of 20µl is recommended for each well of a 96-well plate, which can be adjusted as needed.Note 4: Negative control without DNA template is always recommended.c.Mix well gently by vortex or pipetting. Centrifuge briefly to collect the liquid at the bottom of the PCR tube.2.Transfer qPCR reactions to a qPCR instrument (e.g., ABI QuantStudio™ 6 Flex Real-Time PCR System) and run thermocycling conditions as follows:StepTemperatureDurationCyclesTreatment with UDG Enzyme50ºC5 min1Initial Denaturation95ºC2 min1Denaturation95ºC15 sec40Annealing and Extension60ºC15-30 secMelting Curve (optional)95ºC15 sec160ºC15 sec95ºC15 secNote 1: The qPCR running conditions listed above are for general use only. They can be adjusted based on the template, primer sequence, the length of PCR product or GC content, etc. Note 2: The initial denaturation time is usually 2 min, but can be adjusted to 5 min for the amplification of complex or GC-rich DNA fragments.Note 3: The Taq DNA Polymerase is capable of amplifying at least 300bp in 15 seconds. For amplicons longer than 350bp or with high GC content, it is recommended to increase the extension time to 60 s or to use a three-step method to improve amplification efficiency.Note 4: When the annealing temperature of primers is lower than 60ºC, we recommend using the three-step method for PCR amplification.FAQ:1.Low specificity or low amplification efficiency.a.Primer sequence is not well designed. Use primer design tools to avoid inappropriate GC content, secondary structure, dimer, annealing temperature, length, specificity and other possible problems. Use primers published in the literature or order the tested qPCR primers from SYBR Green qPCR Mix (2X)1/5/25mlD7262-1/5/25ml SYBR Green qPCR Mix (2X, Low ROX)1/5/25mlD7265-1/5/25ml SYBR Green qPCR Mix (2X, High ROX)1/5/25mlD7268S/M SYBR Green One-Step qRT-PCR Kit100/500TD7271-1/5/25ml Probe qPCR Mix (2X)1/5/25mlD7272-1/5/25ml Probe qPCR Mix (2X, Low ROX)1/5/25mlD7273-1/5/25ml Probe qPCR Mix (2X, High ROX)1/5/25mlD7277S/M Probe One-Step qRT-PCR Kit100/500次D7501-1/5/25ml SYBR Green qPCR Mix (2X, UDG)1/5/25mlD7503-1/5/25ml SYBR Green qPCR Mix (2X, Low ROX, UDG)1/5/25mlD7507-1/5/25ml SYBR Green qPCR Mix (2X, High ROX, UDG)1/5/25mlD7509S/M SYBR Green One-Step qRT-PCR Kit (UDG)100/500TD7512-1/5/25ml Probe qPCR Mix (2X, UDG)1/5/25mlD7515-1/5/25ml Probe qPCR Mix (2X, Low ROX, UDG)1/5/25mlD7518-1/5/25ml Probe qPCR Mix (2X, High ROX, UDG)1/5/25mlD7523S/M/L Multiplex Probe qPCR Mix (2X, UDG)100/500/2500TD7528S/M Probe One-Step qRT-PCR Kit (UDG)100/500TFASA011-1pc Sealing Film Scraper1pc/bagFSF002Adhesive Film for qPCR20pcs/packageFSF035-100pcs fluorescent quantitative PCR sealing film (pressure sensitive type)100 pcs/packageFSF039-20pcs fluorescent quantitative PCR sealing film (pressure sensitive type, imported and packaged)20 pcs/packageFSF039-100pcs fluorescent quantitative PCR sealing film (pressure sensitive type, imported and packaged)100 pcs/packageFTUB325-1box qPCR Strip Tubes (0.2ml, flat cap, clear)125 pcs/packageFTUB325-10bxs qPCR Strip Tubes (0.2ml, flat cap, clear)125 pcs/package, 10 pcs/caseFTUB33396-well Plate for qPCR20 pcs/packageFTUB384384-well Plate for qPCR20 pcs/packageFTUB335-1box96-well Plate for qPCR (0.2 ml, without skirt, transparent)10 pcs/packageFTUB335-5bxs96-well Plate for qPCR (0.2 ml, without skirt,10 pcs/package, 5 pcs/caseFTUB337-1box96-well Plate for qPCR (0.2ml, semi-skirted, clear)10 pcs/packageFTUB337-5bxs96-well Plate for qPCR (0.2ml, semi-skirted, clear)10 pcs/package, 5 pcs/case

Specification: 2X, Low ROX, UDG