GC-rich PCR Buffer

GC-rich PCR Buffer Set provides 4 GC-rich PCR Buffers that can be used individually or conjunctly to improve the amplification of DNA fragments with high GC content in PCR.According to relevant literature, it is difficult to use a universal GC-rich PCR Buffer to improve the amplification of all DNA fragments with high GC content. Therefore, aladdin developed this GC-rich PCR Buffer Set for users to optimize the PCR conditions conveniently. Several recommended combinations of GC-rich PCR Buffers are also provided for users to use directly.This product is compatible with a variety of commonly used heat-resistant DNA polymerases, such as Taq DNA polymerase and Pfu DNA polymerase.Precautions：The GC-rich PCR Buffer provided by this kit must be used in conjunction with the magnesium-containing PCR Buffer for a particular DNA polymerase.As PCR reaction is extremely sensitive, any contamination of DNA samples should be avoided. Always set up blank controls without templates.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. The PCR reaction can be carried out by referring to the instructions of the DNA polymerase to be used. Generally, the extension time can be appropriately extended to amplify DNA fragments with high GC content. 2. Set up the PCR reaction in a DNase-free microfuge tube as follows:ComponentFinal ConcentrationVolumeVolumeddH2O or Milli-Q water-(26.75-x)µl(10.7-y)µl10X PCR Buffer (with Mg2+)1X5µl2µl5X GC-rich Buffer1X10µl4µldNTP (2.5mM each)0.2mM each4µl1.6µlTemplate DNA10pg-1µg*xµlyµlPCR Primer Mix (10µM each)0.8µM4µl1.6µlTaq DNA Polymerase (5U/µl)1.25U/50µl0.25µl0.1µlTotal Volume-50µl20µlOr:ComponentFinal ConcentrationVolumeVolumeddH2O or Milli-Q water-(31.75-x)µl(12.7-y)µl10X PCR Buffer (with Mg2+)1X5µl2µl5X GC-rich Buffer0.5X5µl2µldNTP (2.5mM each)0.2mM each4µl1.6µlTemplate DNA10pg-1µg*xµlyµlPCR Primer Mix (10µM each)0.8µM4µl1.6µlTaq DNA Polymerase (5U/µl)1.25U/50µl0.25µl0.1µlTotal Volume-50µl20µl3. More than one GC-rich Buffers can be added in one PCR reaction mix. The preferred GC-rich Buffers and their combinations in a PCR reaction mix are listed as follows:a. 1X GC-rich Buffer Ab. 1X GC-rich Buffer Bc. 0.5X GC-rich Buffer Bd. 1X GC-rich Buffer C e. 2X GC-rich Buffer Cf. 1X GC-rich Buffer C + 1X GC-rich Buffer Dg. 2X GC-rich Buffer C + 1X GC-rich Buffer Dh. 1X GC-rich Buffer C + 0.2X GC-rich Buffer B i. 2X GC-rich Buffer C + 0.2X GC-rich Buffer B 4. According to the preliminary amplification effect, the combination of GC-rich Buffers can be further optimized to determine the best GC-rich PCR Buffer or combination for the amplification of target DNA.