In Vitro Rna Transcription

In vitro Transcription. This section covers a range of products that are designed to produce high yields of high quality RNA. Such applications include synthesis of RNA for nucleic acid amplification, gene expression studies, synthesis of modified RNA transcripts that are resistant to RNAse A-type RNases, studying mechanisms of transcription initiation, rapid, maximum-yield synthesis of RNA.

  • RNA Labeling & Modification

In vitro transcription reaction T7 or SP6 (we use NEB enzymes): PCR product 10 ul (use 1 ug of DNA in DEPC dH20) 10X transcription buffer 2 ul 10X DIG labeling mix 2 ul (Roche, contains DIG-UTP) RNasin (40U/ul) 0.5 ul (Promega, prevents RNase activity) RNA polymerase (15-17U/ul) 1 ul (T7 or SP6) DEPC dH2O 4.5 ul. In vitro transcription is a simple procedure that allows for template-directed synthesis of RNA molecules of any sequence from short oligonucleotides to those of several kilobases in μg to mg. In vitro Transcription In Vitro Transcription Reaction (using Ambion Megascript kit) Note: Wear gloves all the time. Take precaution not to contaminate with RNase – do not touch your face with gloved hands, do not spit in the tubes Important: Make all solutions with DEPC-treated water! 1) Thaw the frozen reagents (or aliquots of).

Random labeled single-stranded RNA probes can be synthesized by in vitro transcription. The reaction is catalyzed by bacteriophage T7 RNA polymerase that incorporates labeled NTPs (mostly UTP) as substitute for their natural counterpart using linear, RNA probe-encoding DNA as template.
The optimal ratio of labeled NTP/NTP in terms of product yield and labeling efficiency depends on both the type of labeled NTP and final application. Individual optimization of labeled NTP/NTP ratio can easily be achieved with the single nucleotide format of our HighYield T7 RNA Labeling Kits. They offer maximum optimization flexibility in contrast to fixed NTP labeling mixes combined with high product yields. The combination of (poly)-HRP-conjugated Biotin/Digoxigenin detection reagents with AF 488 (Alexa Fluor® 488)-, AF546 (Alexa Fluor® 546)- or AF594 (Alexa Fluor® 594)-labeled tyramide further increases detection sensitivity of up to 100-fold.

HaptenFluorophoreCopper-free CLICK
HighYield T7 Digoxigenin RNA Labeling Kit HighYield T7 Fluorescein RNA Labeling Kit HighYield T7 Azide RNA Labeling Kit
HighYield T7 Biotin16 RNA Labeling Kit HighYield T7 Cy3 RNA Labeling Kit
HighYield T7 Biotin11 RNA Labeling Kit HighYield T7 Cy5 RNA Labeling Kit
HighYield T7 AF405 RNA Labeling Kit
HighYield T7 Atto 488 RNA Labeling Kit
HighYield T7 AF488 RNA Labeling Kit
HighYield T7 AF555 RNA Labeling Kit
HighYield T7 AF594 RNA Labeling Kit
HighYield T7 AF647 RNA Labeling Kit

Products & Ordering

HighYield T7 Digoxigenin RNA Labeling KitRNT-101-DIGX
Preparation of randomly Digoxigenin-modified RNA probes by in vitro transcription with Dioxigenin-11-UTP
HighYield T7 Biotin16 RNA Labeling KitRNT-101-BIO16
Preparation of randomly Biotin-modified RNA probes by in vitro transcription with Biotin-16-UTP
HighYield T7 Biotin11 RNA Labeling KitRNT-101-BIOX
Preparation of randomly Biotin-modified RNA probes by in vitro transcription with Biotin-11-UTP

In Vitro Rna Transcription Kit

HighYield T7 Fluorescein RNA Labeling KitRNT-101-FAMX
Preparation of randomly Fluorescein-modified RNA probes by in vitro transcription with Fluorescein-12-UTP
HighYield T7 Cy3 RNA Labeling KitRNT-101-CY3
Preparation of randomly Cy3-modified RNA probes by in vitro transcription with UTP-X-CY3
HighYield T7 Cy5 RNA Labeling KitRNT-101-CY5
Preparation of randomly Cy5-modified RNA probes by in vitro transcription with UTP-X-CY5
HighYield T7 AF405 RNA Labeling KitRNT-101-AF405
Preparation of randomly AF488-modified RNA probes by in vitro transcription with UTP-PEG5-AF405
HighYield T7 Atto488 RNA Labeling KitRNT-101-488
Preparation of randomly Atto488-modified RNA probes by in vitro transcription with UTP-ATTO-488
HighYield T7 AF488 RNA Labeling KitRNT-101-AF488
Preparation of randomly AF488-modified RNA probes by in vitro transcription with UTP-PEG5-AF488
Transcription

In Vitro Rna Transcription Troubleshooting

HighYield T7 AF555 RNA Labeling KitRNT-101-AF555
Preparation of randomly AF555-modified RNA probes by in vitro transcription with UTP-PEG5-AF555
HighYield T7 AF594 RNA Labeling KitRNT-101-AF594
Preparation of randomly AF594-modified RNA probes by in vitro transcription with UTP-PEG5-AF594
HighYield T7 AF647 RNA Labeling KitRNT-101-AF647
Preparation of randomly AF647-modified RNA probes by in vitro transcription with UTP-PEG5-AF647
HighYield T7 Azide RNA Labeling KitRNT-101-AZ
Preparation of randomly Azide-modified RNA probes by in vitro transcription with 5-Azido-C3-UTP
AF488 tyramide reagentRNT-012
also known as Alexa Fluor® 488 tyramide
AF546 tyramide reagentRNT-013
also known as Alexa Fluor® 546 tyramide
AF594 tyramide reagentRNT-014
also known as Alexa Fluor® 594 tyramide

In Vitro Rna Transcription

AF488 Streptavidin ConjugateRNT-015
Streptavidin, Alexa Fluor® 488 Conjugate

Spermidine Rna In Vitro Transcription

AF594 Streptavidin ConjugateRNT-016
Streptavidin, Alexa Fluor® 594 Conjugate