![SMOBIO/[TP1260] ExcelTaq™ 5X Fluorescent PCR Master Mix, 200 Rxn/200 Rxn/TP1260](images/SMOBIO/image.jpg)
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
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抗人球蛋白试验，又称Coombs’试验，是诊断自身免疫性溶血性贫血(AIHA)的重要依据。
正常人直接与间接试验均为阴性，Coombs’试验阴性有时并不能排除AIHA
本试验阳性结果主要见于下列几种情况：
1.自身免疫性贫血，（IgG）型引起的溶血性贫血，本试验直接反应常呈强阳性，间接反应大多阴性，但亦可阳性。
2.药物诱发的免疫性溶血性贫血①α-甲基多巴型：直接及间接反应均阳性。②青霉素型：直接反应阳性，间接反应阴性，以上二型如以正常红细胞先与有关药物于37℃培育后再加病人血清、间接反应均为阳性。③福阿亭型：（奎宁等药物）抗体通常为IgM，偶有IgG型者，直接反应为阳性，间接反应阴性，但如用IgG抗血清做试剂则结果大部分均为阴性，但如培育时加入新鲜正常人血清（供应补体）则结果为阳性。
3.冷凝集素综合征直接反应阳性，间接反应阴性（试验需在37℃下进行）由于本病红细胞膜附着的是补体C4和C3而不是IgG或IgM，如果用抗IgG或抗IgM抗血清做试验时，则结果阴性，如以抗补体的抗血清做试验则直接反应阳性。
4.新生儿同种免疫溶血病，因Rh血型不合所致溶血病，直接及间接反应均强阳性，持续数周、换血输血后数天内可变为阴性，由于“ABO”血型不合引起的溶血病，结果常为阴性或弱阳性。
5.红细胞血型不合引起的输血反应，ABO或Rh血型不合输血，供者的红细胞被受者的血型抗体致敏，在供者被致敏的红细胞完全破坏以前，直接反应阳性，Rh阴性者如过去不曾接受过Rh阳性者的血或曾妊娠胎儿为Rh阳性者，间接反应阳性，如无上述接触，第一次输血后（Rh阳性的血），数天之内间接反应也会变为阳性。
6.其它在传染性单核细胞增多症、SLE、恶性淋巴瘤、慢性淋巴细胞白血病、癌肿、铅中毒、结节性动脉周围炎、EVan氏综合征等，病人直接反应亦可阳性，阵发性寒冷性血红蛋白尿症患者中，急性发作后用抗补体血清做试验直接反应常为阳性。展开
