Navigation Links
Efficient and Reliable PCR Setup Using Eppendorf MasterMix

bjective was to test the reproducibility of the results and to ascertain the stability of the MasterMix at different storage and application temperatures. Tests were carried out on different types of applications and different individual applications.

Materials and Methods

Experiment 1:
Reproducibility of parallel PCR preparations tested using Eppendorf MasterMix
In 56 parallel preparations, a 400-bp fragment from the gene of the Taq polymerase (60% GC content) was amplified. 15 l template/primer mix was added to 10 l of 2.5 x MasterMix (per preparation: 1.5 mM MgCl2, 200 M each of dNTP, 1.25 U Taq; 200 pg pTTQ18 plasmid DNA, 20 pmol per primer). The reaction took place in a 0.2 ml PCR tube with the following PCR program on Mastercycler gradient** from Eppendorf.

94C 3 minute initial denaturation
30 cycles:
94C 15 seconds denaturation
59C 10 seconds annealing
72C 40 seconds extension

The fragments were separated by electrophoresis in a 1.2% agarose gel in TAE buffer at 110 V.

Experiment 2:
Stability test for PCR MasterMix: Using Eppendorf MasterMix after storage under different condition temperatures in three different PCR systems.
Following incubation for 48 hours each at 20C, 28C, 2028C or 37C, Eppendorf MasterMix was tested in three different PCR systems. According to the system used, a 360-bp fragment from the human gene apolipoprotein B (Exon 26), a 547-bp fragment with the coding sequence of the pyrophosphatase gene of Thermo thermophilus (70% GC content) and a 10-kb fragment from the genome of the phage lambda was amplified. The target DNA
'"/>

Source:


Page: All 1 2 3 4 5 6 7

Related biology technology :

1. Efficiently Insert Unique Restriction Sites into Plasmid Vectors
2. Efficient Cleavage of Fusion Proteins to Yield Native Amino Termini
3. Mammalian Expression Vector for Efficient Cloning of PCR Fragments
4. Efficient Transfection of Neurospora Crassa
5. Efficient Transfection of Neurospora Crassa
6. Efficient Recovery of Ultrapure Plasmid DNA
7. Efficient DNA transfection of primary CNS neurons using TransMessenger Transfection Reagent
8. Efficient RNAi-mediated gene silencing in neuronal cells using QIAGEN siRNA and TransMessenger Transfection Reagent*
9. Cloning Based on Efficient Three-Fragment Assembly DNA Ligation
10. Efficient and Reliable Linear Amplification of cRNA
11. Efficient Delivery of siRNAs to Human Primary Cells: Electroporation vs. Chemical Transfection
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Efficient and Reliable PCR Setup Using Eppendorf MasterMix

(Date:9/15/2014)... of carbon nanotubes is neat, but Rice University scientists are ... in new fibers created at Rice line up like a ... Angel Mart and his colleagues. , The tricky bit, according ... the journal ACS Nano , is keeping the densely ... , Left to their own devices, carbon nanotubes form clumps ...
(Date:9/15/2014)... , Sept. 15, 2014 Research and ... Exome Sequencing Market by Product, by Services, by Technology, by ... offering. Whole Exome Sequencing (WES) is next-generation ... genome, i.e. the exon. The exome, which is the total ... the total genome and is actually converted into proteins after ...
(Date:9/15/2014)... septiembre de 2014 La segunda anual International ... 12 al 18 de octubre. Como iniciativa conjunta de la ... compañías miembro s, la IPAW se ha diseñado para: ... a la colección de fuentes de plasma , Reconocimiento ... y mejorar las vidas , Aumento del conocimiento sobre ...
(Date:9/14/2014)... survival strategies. They often create a biofilm to protect ... with antibiotics. A biofilm is a bacterial community that ... sugar chains and "curli". Scientists at VIB and ... a detailed three-dimensional image of the pores through which ... a crucial step in the formation of the protective ...
Breaking Biology Technology:Rice rolls 'neat' nanotube fibers 2Rice rolls 'neat' nanotube fibers 3Global Whole Exome Sequencing (Systems, Kits, Library Preparation, Target Enrichment) Market by Product, by Services, by Technology, by Application - Forecast to 2018 2Global Whole Exome Sequencing (Systems, Kits, Library Preparation, Target Enrichment) Market by Product, by Services, by Technology, by Application - Forecast to 2018 3La International Plasma Awareness Week será una celebración para los donantes 2La International Plasma Awareness Week será una celebración para los donantes 3New insights in survival strategies of bacteria 2
... Corp . is now using radio frequency identification (RFID) ... the world's largest manufacturer of cordless telephones. , ,RFID ... to engage the supply chain management technology, according to ... integrate passive RFID technology into its supply chain environment. ...
... - Alfalight, Inc., announced the release of ... two configurations for delivery in September. , ,The CPM-II is ... (0.22 NA) output fiber for fiber laser applications. A version ... micron signal/pump fiber) is also available for fiber amplifier and ...
... week before Badger football begins its 2005 season, researchers at ... way to demonstrate their school spirit. The only fans who ... , ,Last month a team of UW-Madison chemists led by ... the university's mascot made of nanofiber hairs each only 75 ...
Cached Biology Technology:Nanotechnology minimizes UW's beloved mascot 2
(Date:9/15/2014)... bonds are produced naturally across all kingdoms of life ... a broad range of biological activities. The presence of ... influence on their molecular properties. , One ... to find the late-stage, site-specific incorporation of a halogen ... sp C-H bond (one of the most inert chemical ...
(Date:9/15/2014)... coral reef ecologists fears that reef biodiversity may not ... we once thought. , In an international study published ... Excellence for Coral Reef Studies (Coral CoE) says we ... within reef ecosystems. , In coral reefs, just as ... keep the ecosystem safe and functioning. , Professor ...
(Date:9/15/2014)... assistant professor in the Department of Science and Technology ... Human Sciences at Virginia Tech, has won a 2014 ... study the prospects and problems of creating a global ... Japan,s Fukushima I Nuclear Power Plant in March 2011 ... viewed nuclear emergencies, Schmid said. Three of the plant,s ...
Breaking Biology News(10 mins):Pitt chemical biologist finds new halogenation enzyme 2Specialized species critical for reefs 2'Nuclear disasters don't respect national boundaries' 2
... wrinkles and hide those pesky grays, but what about ... fix, such as losing muscle mass? Cutting calories early ... the phenomenon in rats. A restricted-calorie diet, ... mitochondrial mishap that may contribute to muscle loss in ...
... This press release is available in French . ... Research Institute of the Jewish General Hospital along with ... Research Institute (OHRI) report a significant breakthrough in the use ... known as oncolytic virotherapy. Their results were published in the ...
... to see a single molecule inside a living cell. ... helping to develop a new technique that will enable ... unprecedented look at the atomic structure of cellular molecules. ... the country and in Germany and Sweden to utilize ...
Cached Biology News:Cutting calories could limit muscle wasting in later years 2Cutting calories could limit muscle wasting in later years 3Viral 'magic bullet' targets cancer cells with help of new compound 2Improving our ability to peek inside molecules 2
... your microarray data using Asuragens bioinformatics ... need from consultation to extensive data ... date methods. Utilize our expertise in ... Service or from your own microarray ...
... value of your microarray data using ... support you need from consultation to ... up to date methods. Utilize our ... Expression Profiling Service or from your ...
... Maximize the value of your microarray ... the support you need from consultation to ... to date methods. Utilize our expertise in ... or from your own microarray experiments., ,Normalization ...
... This CLS number is a ... easily match Cornings product number. ... please order under the old ... customer service for assistance. ...
Biology Products: