Description
The ExcelTaq™ 5X/2X 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. The mixture contains all components for PCR with the exception of templates and primers. This not only saves valuable time in the laboratory, but also reduces pipetting and reagent handling errors. The PCR Master Mix is supplied as a 5X/2X concentrated ready-to-use mixture of recombinant Taq DNA Polymerase, reaction buffer, MgCl2 (TP1120 contains MgSO4), dNTP, and enzyme stabilizer enabling efficient amplification of template in PCR and allows the user to prepare a PCR reagent conveniently. This product is supplied with 6X DNA Loading Dye (Blue) containing two tracking dyes (Xylene cyanol FF and Bromophenol blue) for post PCR analysis through the use of agarose gel 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
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
4°C for 6 months-20°C for 24 months
Elongation capability
ExcelTaq™ PCR Master Mix can reliably amplify λDNA up to 8 kb in length. (M: DM3100)
Contents
Component | Volume | Cat. No |
ExcelTaq™ 5X PCR Master Mix | 2 x1 ml | TP1100 (200 Rxn) |
6X DNA Loading Dye (Blue) | 2 x 1 ml | |
ExcelTaq™ 2X PCR Master Mix (MgSO4) | 2 x1.25 ml | TP1120 (100 Rxn) |
6X DNA Loading Dye (Blue) | 1 ml |
Storage
4°C for 6 months-20°C for 24 months
Manual
Manual_TP1100_ExcelTaq™ 5X PCR Master Mix
Manual_TP1120_ExcelTaq™ 2X PCR Master Mix (MgSO4)
SDS
SDS_TP1100
SDS_TP1120
Recommended PCR Condition
|
Recommended PCR Program
Steps | Temp. | Time | Cycles |
Templatedenature | 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 variesaccording to primers’ thermodynamic properties.
Morphological and molecular description of Rhadinorhynchus laterospinosus Amin, Heckmann & Ha, 2011 (Acanthocephala, Rhadinorhynchidae) from marine fish off the Pacific coast of Vietnam.
Amin OM, Heckmann RA, Dallarés S, Constenla M, Ha NV.
Parasite. 2019;26:14. doi: 10.1051/parasite/2019015. Epub 2019 Mar 6.
PMCID: PMC6402367
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.
[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
ebiomall.com
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1 mmol/LEDTA(pH 8.0)
因为含有以上两种物质,所以称为TE。
配制分三步:
1)1 M Tris-HCl (pH 8.0) 50 ml的配制:称取Tris碱6.06 g,加超纯水40 ml溶解,滴加浓HCl约2.1 ml调pH至8.0,定容至50 ml。
2)0.5 M EDTA(pH 8.0)50 ml的配制:称取EDTA-Na2·2H2O 9.306 g,加超纯水35 ml,剧烈搅拌,用约1 g NaOH颗粒调pH至8.0,定容至50 ml。(EDTA二钠盐需加入NaOH将pH调至接近8.0时,才会溶解。)
3)1×TE(10 mM Tris-HCl,pH 8.0;1 mM EDTA,pH 8.0)的配制:
作用:
TE缓冲液是弱碱性,对DNA的碱基有保护性,(DNA在它是的稳定性较好,不易破坏其完整性或产生开环及断裂),包括提取好的DNA也要放在TE缓冲液是保存. 10mMTris-Hcl,pH有7.47.68.0三种。
EDTA调到8.0是为了更好溶解,其他只要调到相应pH就可以。Tris在7-8附近缓冲能力很强,所以加8.0的EDTA下去后,不会改变pH。
碳酸氢钠溶液是一种co2缓冲液,当瓶内二氧化碳量减少时,碳酸氢钠溶液释放co2,反之吸收co2.因此它能保持瓶内二氧化碳量大致不变.
TBST中含有Tris-Hcl,NaCl,Tween20这三种物质,是做WESTERNBLOT中常用的一种缓冲液。
TBST缓冲液的配制
1000ml×TBST的配置
先称量NaCl40g,倒入烧杯中,加DDW蒸馏水400ml,再称量NaCl47.6g,倒入刚才的那个烧杯中(PS:由于NaCl的量太多,一次称量不方便,所以分两次称量,且易于溶解)。往烧杯中加入Tris—HCl缓冲液100ml,最后加(吐温20)5ml,转入1000ml容量瓶中,在定容,转移即可。
TBST缓冲液的应用:
1.主要用于免疫组化和原位杂交,酶联免疫等实验中,清洗免疫印。
2.迹膜;
注意事项:
1.TBST缓冲液,PH7.2-7.5;
2.颜色为无色透明液体;
3.为了您的安全和健康,请穿实验服并戴防护手套操作;
1、 缓冲溶液对反应物的测定没有干扰
2、缓冲组分的浓度为1:1
3、有足够的缓冲容量
4、缓冲溶液的PH应在所需范围内
5、组成缓冲溶液的弱碱PKB和弱酸PKA应接近或等于所需的POH值或PH值(PH+POH=14)
配制
只要知道缓冲对的PH值,和要配制的缓冲液的pH值(及要求的缓冲液总浓度),就能按公式计算[盐]和[酸]的量。这个算法涉及对数换算,较麻烦,前人为减少后人的计算麻烦,已为我们总结出pH值与缓冲液对离子用量的关系并列出了表格。只要我们知道要配制的缓冲液的pH,经查表便可计算出所用缓冲剂的比例和用量。例如配制500nmpH5.8浓度为0.1M磷酸缓冲液。
经查表知pH5.8浓度为0.2M Na2HPO48.0毫升,而0.2M Na2HPO492.0毫升。依此可推论出配制100ml0.1M的磷酸缓冲液需要0.1M Na2HPO48.0毫升,而0.1M Na2HPO4需要92.0毫升。
计算好后,按计算结果准确称好固态化学成分,放于烧杯中,加少量蒸馏水溶解,转移入50ml容量瓶,加蒸馏水至刻度,摇匀,就能得到所需的缓冲液。
各种缓冲溶液的配制,均按表格按比例混合,某些试剂,必须标定配成准确浓度才能进行,如醋酸、氢氧化钠等。另外,所有缓冲溶剂的配制计量都能从以上的算式准确获得。