Synthesis of mRNA - in vitro ..

Image segmentation is a useful step in localizing mRNA expression to specific cell or tissue-types. Oftentimes, it is possible to take advantage of DAPI staining (for demarcating the body axes and major tissue groups, i.e., body wall muscles, ventral nerve cord, intestinal cells, etc.), GFP reporter systems, and smFISH signals (by staining for genes previously known to be expressed in a given cell or tissue) to accurately assign smFISH spots to their tissue of origin. Computational software can facilitate this process by overlaying images from multiple channels (e.g., Cy5, Alexa 594, GFP and DAPI), followed by manual or automatic annotation of the relevant spatial landmarks. These annotations can later be overlaid onto the smFISH images to allow tissue-specific transcript quantification. Additional analysis, such as measurement of tissue length or size, can also be performed computationally at this step. If performing segmentation using computational software (e.g., MATLAB), the resulting traces and annotations can be saved as a standalone data file and used for region-specific spot counting (step 5) later.

Ambion kit: RNA synthesis x µl DEPC treated water

Competitive Quantitative RT-PCR kits (Ambion Inc.) are based on this application.

What are microRNA (miRNA) mimics? – Bio-Synthesis, …

Total RNA, including miRNAs, was polyadenylated andreverse-transcribed with a poly(T) adapter into cDNAs for real-time PCRusing the miRNA-specific forward primer and the sequence complementaryto the poly(T) adapter as the reverse primer.

In Vitro mRNA Synthesis - Heald Lab - Google Sites

To evaluate possiblepre-amplificationbias, we compared the expression of 384 miRNAs in three differentcancer cell lines with Megaplex RT, with or without an additionalpre-amplificationstep.

Additionally, farless input material is required compared to microarrays and otheralternative technologies.

mMESSAGE mMACHINE™ T7 Transcription Kit

Our convenient and high-quality real-time RT-PCR accessory products complete the RT² Profiler™ PCR Array System and the RT² PCR Primer Assays. The other components needed for real-time PCR analysis are SYBR Green PCR master mix, and an optional RNA QC array.

Custom Long RNA Transcription Synthesis Services - …

Real-time RT-PCR-specific errors in the quantification of microRNAtranscripts are easily compounded with any variation in the amount ofstarting material between the samples, e.g.

Order Ambion's Dynabeads® mRNA DIRECT ..

The single molecule fluorescent hybridization (smFISH) method differs from conventional approaches by using many short (about 20 base pairs long) oligonucleotide probes to target different regions of the same mRNA transcript (; ). Each oligo is conjugated with only one fluorophore and thus faintly visible by itself. Binding of multiple oligos to the same transcript yields a bright spot, indicative of a single mRNA transcript. Since mis-bound probes are unlikely to co-localize, this method effectively reduces false-positive signal from non-specific probe binding. The small oligo size allows the probes to efficiently penetrate through target tissue, yielding robust detection of even low-abundant transcripts. Subsequently, the total number of fluorescent spots within a single cell or region can be unambiguously counted and compared across different developmental stages and genetic backgrounds.

Once the packaged RNA is extracted from the protective coat protein, it is completely compatible with reverse transcription.

What is the best method for cDNA synthesis? - …

For this reason, RNA controls and standards are key components of the assays.Currently, these diagnostic assays rely on RNA synthesized by transcription to produce the appropriate controls and standards.

Initiation of Protein S mRNA synthesis in human liver and various ..

My preferred method is to use the Ambion T7 Megascript kit, but I haven't made a full comparison to other kits. They should all be more or less the same. But the transcription itself may not be enough.

and directs degradation of the complementary mRNA sequence by a ..

B. RNA extracted from a FFPE spleen sample were converted to cDNA with or without pre-amplification with RT² PreAmp PCR Master Mix and RT² PreAMP Primer Mix (Custom). The unamplified and PreAMP amplified samples were analyzed on a custom PCR array containing 45 genes. Shown here is raw Ct comparison between unamplified and amplified cDNA. Only genes which have Cts lower than 33 in the unamplified cDNA are shown here.