Description
The ExcelTaq™ 5X Fluorescent PCR Master Mix is a ready-to-use mixture for amplifying targeted DNA fragments. It is designed to serve as a master mix for virtually all PCR applications. This product is supplied as a 5X concentrated mixture containing all the essential ingredients for PCR with the exception of templates and primers. In addition, the mixture contains electrophoresis tracking dye (Bromophenol blue) and a safer fluorescent DNA staining dye, which enables the user to track the electrophoresis process in real time as well as eliminates the need for the staining process. The resultant PCR reaction mixture is sufficiently dense enough to be loaded directly into a TAE or TBE buffered gel for electrophoresis.
Features
5’→3’ DNA polymerase activity
No detectable 3"→5" exonuclease (proofreading) activity
Generates PCR products with 3"-dA overhangs
High yield PCR
High reproducibility
Reduced pipetting errors
For direct loading into electrophoresis analysis
DNA bands can be visualized directly under UV or 470 nm blue light illumination
Applications
Routine PCR
Colony PCR
High throughput PCR
Amplification of DNA fragments up to 8 kb
Generation of PCR products for TA cloning
DNA labeling
Storage
Protected from light and avoid multiple freeze/thaw cycles4°C for 6 months-20°C for 24 months
Elongation capability
λDNA was amplified with ExcelTaq™ 5X Fluorescent PCR Master Mix. The λDNA was amplified with specific primers for amplifying different ranges. The pictures were captured under B-BOX™ blue light as seen in the left picture. While using UV light, the amplicons can be seen in the right picture (M: DM 3160).
Colony PCR
Colony PCR with ExcelTaq™ 5X Fluorescent PCR Master Dye Mix. The JM109 or modified JM109 was used for amplifying the genomic galactosidase Z gene. The pictures were captured under B-BOX™ blue light as seen in the left picture. While using UV light, the amplicons can be seen in the right picture (M: DM 3160).
Contents
Component | Volume |
ExcelTaq™ 5X Fluorescent PCR Master Mix | 2 x 1 ml |
Storage
Protected from light and avoid multiple freeze/thaw cycles4°C for 6 months-20°C for 24 months
Manual
Manual_TP1260_ExcelTaq™ 5X Fluorescent PCR Master Mix
SDS
SDS_TP1260
Why is the size of the PCR product slightly different from prediction when used with TP1260 for PCR reaction?
It is due to variation of the fluorescent dye/DNA ratio. If less DNA was bond with too much fluorescent dye, the DNA would migrate slower in the electrophoresis and presented a larger size than expected. TP1260 therefore is not suitable for running such experiments needed to distinguish subtle differences.
Recommended PCR Condition
|
Recommended PCR Program
Steps | Temp. | Time | Cycles |
Template denature | 94°C | 2 min | 1 |
Denature | 94°C | 30 sec | 25-40 |
Annealing | 50-68°C* | 30 sec | |
Extension | 72°C | 30 sec/kb | |
Final extension | 72°C | 1 min | 1 |
*Optimal PCR condition varies according to primers’ thermodynamic properties.
B-BOX™ Blue Light LED Epi-illuminator
470 nm long wavelength
Improved cloning efficiency
Compact, light-weight, and portable (less than 1 kg)
Adjustable and removable filter plate allows for gel cutting, visualization, and documentation
470 nm long wavelength
Improved cloning efficiency
Compact, light-weight, and portable (less than 1 kg)
Adjustable and removable filter plate allows for gel cutting, visualization, and documentation
FluoroBand™ DNA Ladder series
Sharp bands
Quick reference— enhanced bands
Ready-to-use— premixed with loading dye for direct loading
Stable— room temperature storage over 6 months
Directly observed by UV or blue light— premixed with high sensitive fluorescent dye
FluoroVue™ Nucleic Acid Gel Stain
Excellent for in-gel staining
Sensitivity up to 0.14 ng (DNA) or or 1 ng (total RNA)
A safe alternative to EtBr
Suitable for blue or UV light
Excellent for in-gel staining
Sensitivity up to 0.14 ng (DNA) or or 1 ng (total RNA)
A safe alternative to EtBr
Suitable for blue or UV light
High Fidelity PCR amplification
Amplification of target gene with HiFi™ DNA polymerase to minimize error rate.
[TF1000] SMO-HiFi™ DNA Polymerase, (1 U/μl, 100 U)
[TF3000] G-HiFi™ DNA Polymerase, (1 U/μl, 100 U)
Gel electrophoresis
Staining amplicons with safe fluorescent dyes, following by observation under blue-light illuminator to minimize damage of DNA amplicons and maximize successful cloning efficiency.
Safe fluorescent dyes
[NS1000] FluoroVue™ Nucleic Acid Gel Stain (10,000X), 500 μl
[DS1000] FluoroStain™ DNA Fluorescent Staining Dye (Green, 10,000X), 500 μl
[DL5000] FluoroDye™ DNA Fluorescent Loading Dye (Green, 6X), 1 ml
Blue-light illuminator
[VE0100] B-BOX™ Blue Light LED Epi-illuminator
Ligation
Blund-end PCR amplicons can directly ligate with PCR cloning vector.
[CV1000] GetClone™ PCR Cloning Vector, 20 Rxn
[CV1100] GetClone™ PCR Cloning Vector II, 20 Rxn
Transformation
Prepare competent cells with high efficiency and transform with time-saving protocol.
[CK1000] Champion™ E. coli Transformation Kit
