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:6/2/2015)... 2015 /CNW/ - Shell today celebrated the North American launch ... with an exhibit the size of a city block ... for media and the public, and special appearances by ... Gasoline provides the best total engine protection by addressing ... gunk and corrosion. "With the launch of ...
(Date:6/2/2015)... , June 02, 2015 Research and ... "Molecular Diagnostics - Global Market Size, Strategy and ... offering. "Molecular Diagnostics - Global Market Size, ... that analysts and planners can use. Hundreds of pages ... United States Medicare Fee Payment Schedules to help understand ...
(Date:6/2/2015)... PAULO , June 2, 2015   Hospira, ... in biosimilars, today announced that its partner, Celltrion – a ... – the National Health Surveillance Agency in ... biosimilar monoclonal antibody approved for use in ... Hospira. Hospira also markets and sells biosimilar infliximab in ...
(Date:6/2/2015)... , June 2, 2015   Sigma-Aldrich Corporation ... Science and Technology company, announced today it has ... to launch genetically engineered human neural differentiated ... the research of neurological diseases. These new products, ... unique and versatile tools for the study of ...
Breaking Biology Technology:New Shell V-Power NiTRO+ Premium Gasoline Introduces Breakthrough Formulation for BEST TOTAL Engine Protection 2New Shell V-Power NiTRO+ Premium Gasoline Introduces Breakthrough Formulation for BEST TOTAL Engine Protection 3New Shell V-Power NiTRO+ Premium Gasoline Introduces Breakthrough Formulation for BEST TOTAL Engine Protection 4Global Molecular Diagnostics Report 2015-2019 - Market Size, Strategy and Forecasts 2Global Molecular Diagnostics Report 2015-2019 - Market Size, Strategy and Forecasts 3ANVISA Approves First Infliximab Biosimilar in Brazil 2ANVISA Approves First Infliximab Biosimilar in Brazil 3ANVISA Approves First Infliximab Biosimilar in Brazil 4ANVISA Approves First Infliximab Biosimilar in Brazil 5Sigma-Aldrich to Launch XCell Science Genetically Engineered iPSC-Derived Neural Disease Cells 2Sigma-Aldrich to Launch XCell Science Genetically Engineered iPSC-Derived Neural Disease Cells 3
... MADISON - Berbee Information Networks Corporation s recent ... geographic expansion, but it also brings NEAs strength in ... into Berbees portfolio. , ,Berbee announced it bought the ... and that it expected to keep all of NEAs ...
... MILWAUKEE A new technology at Childrens Hospital of ... irregular beats, in children by freezing the tissue causing ... CryoCath Technologies reaches temperatures as low as ... the catheterization laboratory at Childrens Hospital, an electrophysiologist and ...
... , ,Are your projects still late? Do they continue to ... the answer is probably yes. , ,The point of this ... Think about it you learn new things every day. You ... gone to class. Youve learned the latest techniques. The same ...
Cached Biology Technology:Berbee acquisition could expand VoIP in Wisconsin 2Project Performance: Is your project late? Over budget? 2Project Performance: Is your project late? Over budget? 3
(Date:5/11/2015)...  Through a well-rounded UAS delegation representing private industry, government, ... showing at AUVSI,s Unmanned 2015 conference last week in ... UAS industry met with over 200 hundred people ... the UAS ecosystem. "Our message is clear ... Aerospace Rich Knoll . "If you want to fly ...
(Date:5/6/2015)... , May 6, 2015 LifeBEAM, a developer ... helmet producer, announced today that they will expand their ... world,s first bio-sensing cycling helmet and the first joint ... two new colors in order to give cyclists more ... In addition, LifeBEAM and Lazer announced their plan to ...
(Date:4/21/2015)... , April 21, 2015 ... government policies are boosting access control systems market in ... According to a recently published report by TechSci Research ... ", the access control systems market in ... billion by 2020.The access control systems market in the ...
Breaking Biology News(10 mins):Ohio Flies High at Unmanned '15, Sets Stage for Ohio UAS Conference 2LifeBEAM and Lazer Announce Expanded Partnership in Bio-Sensing Sports Gear 2LifeBEAM and Lazer Announce Expanded Partnership in Bio-Sensing Sports Gear 3Saudi Arabia Access Control Systems Market to Touch USD1.2 Billion by 2020 Says TechSci Research 2Saudi Arabia Access Control Systems Market to Touch USD1.2 Billion by 2020 Says TechSci Research 3
... Institutet have identified an important mechanism that regulates how ... cell. The study is published in the latest issue ... help us develop new drugs for things like neurological ... most organs of the body, old cells are continually ...
... To a tiny tadpole, life boils down to two basic ... The more a tadpole eats, the faster it grows big ... being active, which ups the odds of becoming someone else's ... activity levels in response to predation risk, but new research ...
... discovered how the brain creates a scent symphony from signals ... researchers found that nerve cells in the brain's olfactory bulb ... do not perceive complex scent mixtures as single objects, such ... cells, or neurons, detect the host of chemical compounds that ...
Cached Biology News:New stem-cell findings can help the body to cure itself 2Time of day tempers tadpoles' response to predators 2Researchers show how brain decodes complex smells 2
... White crystalline solid. HYGROSCOPIC. Water-soluble ... (Cat. No. 252005) but differing in the ... for the extraction of mycoplasma membrane antigens ... ≥98% by TLC. Soluble in H ...
Goat polyclonal to STMN2 ( Abpromise for all tested applications). Antigen: Synthetic peptide: AKTAMAYKEK, corresponding to C terminal amino acids 2-11 of STMN2. Entrez Gene ID: 11075 Swi...
Anti-Human alpha2-Antiplasmin Purified (Polyclonal) (goat IgG)...
... proud to work in collaboration with Pall ... syringe filters in a variety of membrane ... for use on the Caliper TPWII, MultiDose, ... Pricing for our Automation Certified Syringe Filters ...
Biology Products: