The primer is allowed to anneal to the RNA and reverse transcriptase is used to synthesize cDNA from the RNA until it reaches the 5' end of the RNA. (wikipedia.org)
By denaturing the hybrid and using the extended primer cDNA as a marker on an electrophoretic gel, it is possible to determine the transcriptional start site. (wikipedia.org)
These applications include next-generation sequencing, transfection, real-time PCR, measuring yields of RNA preparations for microarray experiments, reverse transcription PCR (RT-PCR), differential display PCR, northern blot analysis, S1 nuclease assays, RNase protection assays, and cDNA library preparations. (thermofisher.com)
The RiboGreen reagent is ideal for fast and accurate measurement of RNA before generating cDNA, northern blot analysis, S1 nuclease assays, RNase protection assays, reverse transcription PCR, and differential display PCR. (thermofisher.com)
The recommended final concentration for RNA protection is about 1 -2 units of RNase Inhibitor for every 1 µl of the reaction mixture. (biotechrabbit.com)
RNA synthesized from this kit is suitable for many applications including RNA structure and function studies, ribozyme biochemistry, probes for RNase protection or gel shift assays, hybridization-based blots, anti-sense RNA or RNAi experiments, microarray analysis, microinjection, sgRNA synthesis and in vitro translation studies. (neb.com)
Detection Assays1
The surging COVID19 pandemic has underlined the need for quick, sensitive, and high-throughput SARS-CoV-2 detection assays. (medrxiv.org)
Genomic2
OliGreen reagent assays are routinely used for antisense oligonucleotides, aptamers, genomic DNA isolated under denaturing conditions, PCR primers, sequencing primers, and single-stranded phage DNA. (thermofisher.com)
performed on genomic DNA, using a 50 nuclease PCR assay. (cdc.gov)
Quantitation2
PicoGreen reagent assays provide a rapid and simple fluorescence-based method for the accurate quantitation of dsDNA. (thermofisher.com)
The assay is many times more sensitive than conventional UV absorbance, and free nucleotides or short oligonucleotides of up to 6 bases do not interfere with the OliGreen ssDNA quantitation assay. (thermofisher.com)
Probes1
Endogenous control is to be used with Inventoried and Non-Inventoried TaqMan™ Gene Expression Assays, Custom TaqMan™ Gene Expression Assays, and⁄or Custom TaqMan™ Primers and Probes. (fishersci.com)
Vitro3
These include S1 nuclease protection and primer extension ( Chang and Clayton 1984 ), as well as in-vitro capping ( Yoza and Bogenhagen 1984 ). (biorxiv.org)
The TPP1/POT1 heterodimer also promotes telomerase processivity, as demonstrated by an in vitro direct telomerase activity assay [ 4 , 20 ]. (prolekarniky.cz)
Cell-based and in vitro assays show that Retro-2 blocks delivery of newly-synthesized TA-proteins to the ER-targeting factor ASNA1 (TRC40). (elifesciences.org)
Extension11
Primer extension is a technique whereby the 5' ends of RNA can be mapped - that is, they can be sequenced and properly identified. (wikipedia.org)
Primer extension can be used to determine the start site of transcription (the end site cannot be determined by this method) by which its sequence is known. (wikipedia.org)
Primer extension offers an alternative to a nuclease protection assay (S1 nuclease mapping) for quantifying and mapping RNA transcripts. (wikipedia.org)
The hybridization probe for primer extension is a synthesized oligonucleotide, whereas S1 mapping requires isolation of a DNA fragment. (wikipedia.org)
Unlike S1 mapping, however, primer extension can only be used to locate the 5'-end of an mRNA transcript because the DNA synthesis required for the assay relies on reverse transcriptase (only polymerizes in the 5' → 3' direction). (wikipedia.org)
Primer extension is unaffected by splice sites and is thus preferable in situations where intervening splice sites prevent S1 mapping. (wikipedia.org)
Finally, primer extension is more accurate than S1 mapping because the S1 nuclease used in S1 mapping can "nibble off" ends of the RNA-DNA hybrid or fail to degrade the single-stranded regions completely, making a transcript either appear shorter or longer. (wikipedia.org)
https://www.nationaldiagnostics.com/electrophoresis/article/primer-extension Shenk, T. E., C. Rhodes, P. W. J. Rigby, and P. Berg. (wikipedia.org)
Here we report on a simple and highly accessible approach to convert commercially available customized DNA microarrays of any complexity and density to RNA microarrays via a T7 RNA polymerase-mediated extension of photocrosslinked methyl RNA primers and subsequent degradation of the DNA templates. (nature.com)
After the DNA library is sequenced on the Illumina flow cell, it is enzymatically double stranded by extension of a 5′ biotinylated primer. (nature.com)
Notably, mutations in the TPP1 OB-fold domain compromise telomerase-dependent telomere extension but not telomere end protection [ 18 , 21 ], indicating that TPP1 performs a role in telomerase regulation which is distinct from its contribution to chromosome end protection. (prolekarniky.cz)
Reagent2
PicoGreen reagent assays are routinely used for genotyping by allele-specific PCR, quantitating dsDNA samples before and after PCR amplification, and determining PCR amplification yields before sequencing. (thermofisher.com)
DNA footprinting utilizes a DNA damaging agent (either a chemical reagent or a nuclease) which cleaves DNA at every base pair. (lookformedical.com)
Electrophoresis1
Specific primers sequences for detection of Topoisomerase II (gyrA) genes were used by conventional PCR then, PCR product were treated with Sac II enzyme at 37°C for 2 h, then restriction bands were recovered by gel electrophoresis. (microbiologyjournal.org)
Gene expression2
Other techniques include dot blotting, ribonuclease protection assays (RPAs), serial analysis of gene expression (SAGE), and differential or subtractive hybridization. (idtdna.com)
Eliminate months of assay design, formulation, and testing by using TaqMan™ Endogenous Controls as your controls to quantify gene expression. (fishersci.com)
Sequence1
There is a random sequence of usually 40 nucleotides (nt) that is flanked by two PCR primer recognition sequences. (neoaptamers.com)
Probe3
This product uses well characterised primers and probe for the N gene. (ha-tech-ltd.com)
Probe is labeled with 6FAM™ dye - MGB and the primers are not limited. (fishersci.com)
This convenient collection of pre-designed probe and primer sets enables you to normalize the amount of sample RNA or DNA in a reaction. (fishersci.com)
Double stranded2
We suggest using Invitrogen's Qubit fluorometric assay , which is specific for double-stranded DNA. (harvard.edu)
The mammalian TRF1 and TRF2 proteins, for example, bind double-stranded telomeres via a Myb-like DNA-binding domain and are involved with telomere length regulation and chromosome end protection. (biomedcentral.com)
Concentration1
The assays are many times more sensitive than conventional UV absorbance and can be optimized for different concentration ranges. (thermofisher.com)
Chromosome end protection1
In this study, we have identified TIN2-R282H, the most common DC-causing mutation in shelterin subunit TIN2, as a separation-of-function mutant which impairs telomerase recruitment to telomeres, but not chromosome end protection. (prolekarniky.cz)
Protocol1
An extraction-less validation protocol was carried out to determine performance characteristics of the assay in both synthetic SARS-CoV-2 RNA as well as clinical specimens. (medrxiv.org)
Tubes1
DEPC-treat all tubes and pipette tips or use certified nuclease-free labware with aerosol filters. (biotechrabbit.com)
Subsequent1
Shortening of the 5'-terminus of the daughter strand, caused by the removal of the terminal RNA-primer and the subsequent incomplete replication of linear DNA molecules, is observed during the genome replication occurring upon cell fission. (actanaturae.ru)
Specific3
Oligo (dT) or gene-specific Primer incubate at 50-55°C for 20-60 minutes. (biotechrabbit.com)
Please do not modify your target-specific PCR primers by adding any universal tags! (harvard.edu)
Specific protein-binding measures are often used as assays in diagnostic assessments. (lookformedical.com)
Activity1
Our antibacterial and antifungal activity assay showed that four fungal species and nine bacterial species have the potential to suppress the growth of at least one seed-borne pathogen tested in the study. (ppjonline.org)
Analysis1
Familiarize yourself with critical parameters of qPCR assay design, specificity analysis, data interpretation, and troubleshooting. (idtdna.com)
Design1
Order ready-to-use PrimeTime™ qPCR Assays or you can design your own custom assay. (idtdna.com)
Provide1
For therapeutic applications you need to use modified nucleotides at all positions in the selection process and this will result in lower synthesis costs on your final commercial aptamer than with a mixture of nucleotides and will provide the level of nuclease protection necessary. (neoaptamers.com)
Control2
No template control (Nuclease free water) checks for cross contamination of samples and contamination of reagents. (ha-tech-ltd.com)
We build each endogenous control using our proven 5' nuclease chemistry. (fishersci.com)
Template1
Sanger sequencing), preferably by using the same primer on the DNA template strand. (wikipedia.org)
Chemical1
There are several strategies that have been proposed to tackle this problem, such as adding nuclease inhibitors or chemical modification of linear DNA ends for protection. (jove.com)
Research2
Massively parallel experimental assays, particularly DNA microarrays and sequencing, have revolutionized genomics research in the last decades. (nature.com)
Integrated DNA Technologies acquires Archer™ next generation sequencing research assays to advance actionable scientific discoveries. (idtdna.com)
Methods1
Although many different methods to detect SARS-CoV-2 particles in clinical material have been developed, none of these assays are successful in combining all three of the above characteristics into a single, easy-to-use method that is suitable for large-scale use. (medrxiv.org)
Requires1
This technique requires a radiolabelled primer (usually 20 - 50 nucleotides in length) which is complementary to a region near the 3' end of the mRNA. (wikipedia.org)