long

Brilliant Core Reagent Kits for QPCR and QRT-PCR

... transcriptase has produced high yieldsof cDNA and long transcripts, providing ideal cDNA templates for subsequent PCRamplifications. 4 The Brilliant single-step QRT-PCR core reagent kits include all the necessaryreagents for cDNA synthesis and PCR amplification in a single tube. This formatuses primers...

Safe Production of High-Titer Retrovirus

...e of interest inserted into a pair of 600-bp viral long terminal repeats (LTRs). The LTR is divided into three regions. The U3 region, containing the retroviral promoter/enhancer, is flanked in the 3 direction by the R region, which contains the viral poly-adenylation signal ...

Pfu DNA Polymerase as a Hot Start Alternative

...enesis; (2)can be used alone to amplify relatively long PCR targets (up to 12 to 25 kb) ifgiven adequate extension time, 12 which is important if long genesequences or vector constructs must be correctly amplified for further sequenceanalysis; (3) unu...

Comparing Fidelity and Performance of Proofreading PCR Enzymes

... DNA polymerase. As described , amplification of long genomic targets (>9 kb) by PfuTurbo DNA polymerase is limited by buffer components, rather than the robustness of the enzyme. Increasing dNTPs (to 500 M) and PCR buffer concentration (to 1.5X) was sufficient to allow PfuTu...

PCR Performance Comparisons Between pfuturbo and Taq DNA Polymerases

...ymerase had difficulty amplifying these relatively long amplicons (lanes11 and 12). Fidelity Comparisons The error rates of a number of DNA polymerases have been measured previouslyin a PCR-based forward mutation assay, which utilizes the lacIOlacZ a target gene. 3 In these studies, Pfu DNA polymerasee...

prostar RT-PCR Systems for Robust High-Fidelity RNA Amplification

...Figure 1 , Panels A and B). In addition to long target capability, the ProSTAR Ultra HF RT...ts (300 to 550 bps; Figure 3, Panel A) and long targets from low-abundance message (1.8- to 5.9-kb dystrophin fragments; Figure 3, P...

Performance Comparisons of Commercial RT-PCR Systems

...gets in high yield, including a relatively long amplicon that was 7.6 kb in length (Panel C). The ThermoScript RT-PCR system with Platinum Taq DNA Polymerase High Fidelity also amplified all three targets. However, product yields were consistently higher with the ProSTAR ...

High-Efficiency Site-Directed Mutagenesis of Large Plasmid Vectors

...nesis kit to improvethe efficiency of mutagenizing long or difficult-to-mutagenize plasmid DNAtemplates using this single day mutagenesis method. The QuikChange XL kitfeatures components specifically designed for more efficient DNA replication andbacterial transformation, which are important when mutageni...

New Protocols for Isolating High- Molecular-Weight Genomic DNA

....** ,4 Because the shuttle vector is nearly 50-kb long and ispresent in multiple copies within the genome of the animal, efficient packagingis contingent on genomic DNA preparations of very high-molecular weight. 6 The DNA must also have sufficient purity so as to be free of substances thatinhibit the pa...

Simple, Sensitive Isotyping of Mouse Monoclonal Antibodies

... card. No agitation, mixing of reagents or long incubation periods are necessary. The IsoDetect card is completely self-contained and is ideally formatted for archiving permanent records. For added convenience, Stratagene is also offering the IsoDetect isotyping control kit...

Easily Amplify Genomic DNA with Long-Distance PCR

...cus. For example, when using targeting vectorswith long arms, 5 conventional PCR is not suitable for mousegenotyping. 6 Hence, we developed a procedure to amplify large DNAfragments with TaqPlus Long DNA polymerase using genomic DNA from mouseear clips. Genomic DNA Prepared from Mouse Ear Clips The in...

Efficient Cleavage of Fusion Proteins to Yield Native Amino Termini

...om recombinant EKdigests without buffer changes or long incubation times. REFERENCES LaVallie, E.R., et al. (1983) J. Biol. Chem. 268: 2331123317. CollinsRacie, L.A., et al. (1995) Biotechnology 13: 982987. Zheng, C.F., and Simcox, T.G. (1996) Strategies 9: 2021. Wybor...

Randomize Gene Sequences with New PCR Mutagenesis Kit

...hod is robust and can be applied to mutagenesis of long targets (1-10 kb). Error-prone PCR is a random mutagenesis technique for introducing amino acid changes into proteins. Mutations are deliberately introduced during PCR through the use of error-prone DNA ...

Unique DNA Polymerase Formulation Excels in a Broad Range of PCR Applications

... polymerase blends areused to successfully amplify long targets (>5 kb), and additionally mayprovide h... to reducedproduct yield and poor amplification of long targets. Moreover, DNA polymeraseblends and PCR buffers optimized for amplification of long targets ...

Enhanced Amplification of Long Targets with PfuTurbo DNA Polymerase

...Holly Hogrefe Stratagene Improve amplification of long targets with PfuTurbo DNA polymerase* by making s... been developed to furtherimprove amplification of long genomic targets with PfuTurbo DNApolymerase. We found that amplification of complex targets greater ...

High-Fidelity PCR with a Novel Polymerase Mixture

... competitors Taq DNA polymerase preparations using long (8minute) extension times. Pfu DNA Polymerase for...PCR system was developed for PCR amplifications of long DNAtarget regions (up to 35 kb) using relatively short extension times (0.5-1minute per kb target). ...

High-Titer Retroviral Vectors for Gene Delivery

...backbone containing a pair of 600-bp viral long terminal repeats (LTRs) between which the gene of interest is inserted. The LTR is divided into three regions: The U3 region contains the retroviral promoter/enhancer, and is flanked in the 3 direction by the R region, ...

Imaging Chemiluminescent Western Blots

...rated density measurements of bands. 1 As long as pixels within analysis areas are not saturated and the dimensions of the analysis areas are kept constant, band concentration is directly proportional to integrated density. Consequently, under similar experimental conditio...

Optimizing pfuturbo DNA Polymerase Amplification Reactions with Perfect Match PCR Enhancer

...pt we employed a 60Cannealing temperature. For the long targets, the amplified reaction productswere elect... enhancer must be present to amplify relatively long targets (e.g., Figure 1 and Figure 2), it may act by reducing secondary structure fo...

Unique Enhanced DNA Polymerase Delivers High Fidelity and Great PCR Performance

...mulated specifically for high-fidelity and long PCR applications. 1 Fig.7 Conclusions As demonstrated here and previously, 1 Herculase enhanced DNApolymerase excels in a broad range of PCR applications. Herculase DNA polymerasecan be readily used in...

Sequence-Validated and Expression-Tested Human cDNA in a Dual Expression Vector

... GFP was chosen because it is easily detectedunder long wavelength UV in induced plate and liquid cultures and requires theformation of a homodimer for biological activity. Results of the enzymatic GFPassays demonstrate that GFP tagged with c-myc and HIS6 was biologically active;therefore, the presence of...

Fast Isolation of Total RNA from Small Numbers of Cells

...by successful RT-PCR amplification of a relatively long target. Primers specific for replication factor C (RFC) were used in RT-PCR ( Figure 3 ) and the expected 3.1-kb band was observed. It has been reported that RFC RNA undergoes alternative processing, which leads to mRN...

Sample Preparation for Microinjection

...remain in the medium especially during long experiments when large numbers of cells are injected. In order that the open petri dish has a sufficient pH stability under the microscope, a carbonate-fr...

TripleMaster PCR System

... 11.9 Primer sequences for long targets ... Tuning Buffer is designed for high fidelity long range PCR applications resulting in robust amplification of genomic targets >20 kb and epi...

Principles and applications of electrical cell fusion

...ng mammals from embryonic cells was carried out as long ago as the late 1980s [6 -10] and aroused little public interest at the time. Vaccination against cancer The results of Vaccination against cancer [11], which have rece...

Principles and applications of electrical cell fusion

...ng mammals from embryonic cells was carried out as long ago as the late 1980s [6 -10] and aroused little public interest at the time. Vaccination against cancer The results of Vaccination against cancer [11], which have rece...

TripleMaster PCR System

... 11.9 Primer sequences for long targets ... Tuning Buffer is designed for high fidelity long range PCR applications resulting in robust amplification of genomic targets >20 kb and epi...

cMaster RTplusPCR System for increased sensitivity and product length in RT-PCR

... factor in One Step applications. Performing long range RT-PCR in a One Step protocol is no longer a...R Kit, special One Step RT-PCR protocol for long templates. Set-up for One Step RT-PCR reactions ...

Efficient and Reliable PCR Setup Using Eppendorf MasterMix

...10 kb). Specific, high-yield amplification of very long fragments (10 kb) is also possible using MasterMix without the need to purchase special additional long-range enzymes. Safe to use and with optimized time requirements, MasterMix is ideal for high-throughput laboratories. ...

HotMasterAn innovative Hot Start/Cold Stop technology for better PCR* results

...tion, which impart disadvantages such as a long initial denaturing step or excessive protein ...on, they impart some disadvantages such as long pre-incubations at high temperatures or protein contamination. Eppendorf has develop...

The use of TaqMaster PCR Enhancer

... DNA isolation procedures from tail slices involve long incubations (one hour to overnight) at high temperature in the presence of Proteinase K. One of the problems that researchers often see is inconsistency in the amplification of transgenic DNA caused by impurities in the templa...

Optimizing DNA Amplification Protocols using the Eppendorf Mastercycler

...tarity within a primer by virtue of palindromes or long segments of polypurines and polypyrimides,...conds are usually fully adequate (4). Very long annealing times normally do not improve yield, but rather produce an increase in spurious pr...

7 Programming

...ith Long PCR, this function can be used to amplify long fragments for a successive extension of the elongation. In the example given, the elongation time is extended by 10 seconds for each cycle, which means that, for the 30th cycle, the elongation time is 290 seconds ...

The TripleMaster PCR System for three different amplifications

... However, demanding applications (such as long range PCR [1-2], high-fidelity PCR, or the a...nd two innovative buffers (Tuning Buffer for long range PCR and High Fidelity Buffer for short range PCR), the TripleMaster PCR System produces ...

TripleMaster and Perfect gDNA Blood Mini Kit Team Up to Amplify Long gDNA Templates

... and Eppendorf TripleMaster PCR System to amplify long templates. In the past, scientists needing ...er team up to enable easy amplification of long templates. Materials and Methods Pulse Field Gel Electrophoresis: A number...

TripleMaster PCR SystemThe Ideal Choice for Long-Range PCR

...o the inability of standard PCR to amplify long segments of DNA is template degradation during exp... effect on long-range PCR is more profound because long templates simply have more sites that are susceptible to depurination. In addition, ...

The QIAGEN Guide to Animal Cell Culture

... cause turbidity, they can go undetected for long periods of time and can easily spread to other cell cultures. The negative effects of mycoplasmal contamination include inhibition of metabolism and growth, as well as interference with nucleic acid synthesis and cell antigenicity. ...

QIAGEN Multiplex PCR Handbook

...tration of 0.2 M. For amplification of long targets (>= 1.5 kb), a primer concentration of... 12. PCR product too long The optimized protocols allow amplification of target sequences up to 1.5 kb. W...

HotStarTaq PCR Handbook

... 15. PCR of long fragments from genomic DNA ...he protocol for amplification of long PCR products using ProofStart DNA Polymerase and QIAGEN Taq DNA Polymerase (see th...

TransMessenger Transfection Reagent Handbook

...sing cells with temperature shifts and long periods without medium during washing steps. It is particularly important for RNA transfection that the cells are in good condition. Therefore, ensure that cell density is not too low at transfection. For adherent ...

Transfection Reagent Selector Kit Handbook

...sing cells with temperature shifts and long periods without medium during washing steps. We recommend performing transfection experiments in the presence of serum, so that cells are not deprived of necessary growth factors and nutrients. ...

FastStart Taq DNA Polymerase

...ication of genomic DNA and cDNA targets up to 3 kb long with high specificity, sensitivity, and yield Multiplex PCR Difficult templates e.g. secondary structures or GC-rich sequences Carry-over prevention Automated PCR e.g. handling at room tem...

Product overview

... Expand Long Template PCR System simplifies long and accurate PCR reactions. This optimized e...duced. Even if Taq were able to amplify such long fragments, Expand Long Template would deliver twice the fidelity and much higher yields. ...

Expand 20 kbPLUS PCR System

for amplification of long targets (up to 35 kb) from eukaryotic genomes ... Application Produces extra long PCR products from genomic DNA. The Expand 20 kb PLUS PCR System is ideal for amplifying ...

Expand Long Template PCR System

... Application Produces a high yield of long PCR products from episomal and genomic DNA. ...da-phage Amplifying full-length cDNAs or long fragments of cDNA (RACE) Expand Reverse Transcriptase is a genetically engineered vers...

High Fidelity PCR Master

...because High Fidelity PCR Master will amplify even long templates without requiring adjustment of reaction buffer components Works with low amounts of template, because High Fidelity Master is more sensitive than other DNA polymerases Convenient storage at 4C means ...

PCR Core KitPLUS

... lead to degradation of PCR product. For long term storage, PCR product should therefore be frozen at 20C. Key advantages Convenient, includes all necessary reagents (except template) to perform PCR and carry-over preve...

Uracil-DNA Glycosylase

... lead to degradation of PCR product. For long term storage, PCR product should therefore be frozen at 20C. Key advantages Prevents carry-over contamination in PCR, because the enzyme modifies all uracil-contain...

PCR reaction components

... For most routine PCR, Taq DNA Polymerase has long been the standard PCR enzyme. However, Taq DNA Polymerase has its limitations. Our Product Selection Guide will provide more information on choosing the appropriate DNA polymerase. Here are some general guidelines: ...

RT-PCR procedures

... Amplifies long sequences. With the right combination of reverse transcriptase and thermostable DNA polymerase, two-step RT-PCR can amplify RNA sequences up to 14 kb long. 2. One tube, one-step procedure...

Properties of PCR Enzymes

...d to produce enzyme blends that could amplify very long DNA (up to 35 kb targets from a genomic template; see Expand Long Template PCR System and Expand 20 kbPLUS PCR System ). Finally, as the demand for more robust PCR, higher yield and fidelity increased, blends of two pol...

Thermal Cycling Profile for Standard PCR

...mplete denaturation of template DNA. Unnecessarily long denaturation times decrease the activity of Taq DNA Polymerase. Primer annealing For most purposes, annealing temperature has to be optimized empirically. The choice of the primer annealing temperature i...

SAGE & LONGSAGE SEQUENCING

...uencing with industry-leading quality. Our long Phred20 read lengths and high sequencing pass rates allow us to obtain high quality sequence data for approximately 40 SAGE tags or 35 LongSAGE tags per read. PROVEN SAGE EXPERIENCE Agencourts proven experience...

Transcriptor Reverse Transcriptase

... making it the product of choice for very long eukaryotic mRNA (Figure 4). To achieve h...PCR System is recommended for the amplification of long fragments (> 5 kb). Result: Trans...

Increase the Power and Sensitivity for Your cDNA Synthesis with the New Transcriptor Reverse Transcriptase

...product ofchoice for reverse transcription of very long eukaryoticmRNA, as shown in Figure 4 for human dystrophin, oneof the longest mRNAs with a length of 14 kb. To achievehigh sensitivity and yield in the following PCR, useFastStart Taq DNA Polymerase or the new FastStart HighFidelity Enzyme Blend. Both...

Increased Sensitivity in Microarray Analysis

...it) is optimized for the efficienttranscription of long mRNA, resulting in high yieldsand sensitivity of cRNA targets. For a direct comparison of sensitivity and reproducibilityof target preparation, we have generated target triplicatesfrom two different human leukemia research s...

The DIG System Nonradioactive and Highly Sensitive Detection of Nucleic Acids

...Film) or with an imaginginstrument. Because of the long lasting glow, multipleexposures are easily achieved. Blots developed withCSPD or CDP-Star can be easily stripped and reprobedif nylon membranes are used. NBT/BCIP The advantage of using a color substrate such...

siRNA Design Guidelines

...can be readily cloned into a vector for long term studies. Ambion scientists have determined that sequences that function well as transfected siRNAs also function well as siRNAs that are expressed in vivo. The only exception is that siRNA sequences to be express...

Determining the Signal-to-Noise Ratio and Optimal Photomultiplier gain setting in the GenePix 4000B

...s is anywhere in the range from 500 V to 900 V, as long as the brightest signals are below saturation. Further reading Basarsky, T., Verdnik, D. Zhai, J.Y. and Wellis, D. Overview of a Microarray Scanner: Design Essentials for an Integrated Acquisition and An...

A Better Way to Isolate RNA AND Protein From the Same Sample

...ue collection, to stabilize RNA during long period of storage, or to circumvent the need to grind frozen tissue into a powder prior to RNA isolation. (See the article " RNArticles: Keeping RNA Intact Prior to RNA Isolation - RNAlater " for some o...