All things RNA: amplify, sequence, ligate, label
Detect and manipulate RNA through amplification
Reverse transcriptases are enzymes able to polymerize a strand of DNA (cDNA) that is complementary to an original RNA template. RNA is susceptible to degradation by RNases and using a reverse transcriptase enzyme to produce cDNA overcomes the problems of working with mRNA. The cDNA becomes the stable template in a variety of downstream applications for RNA studies such as the analysis of gene expression. Regular PCR, qPCR, one-step RT-qPCR or isothermal methods can be used for amplification of the cDNA template. Amplification can be followed by cloning with conventional enzyme protocols or by ligation-independent cloning utilizing the 3’→5’ exonuclease activity of T4 DNA polymerase.
All reverse transcriptase enzymes (RNA-dependent DNA polymerases) can be used to:
Choose a reverse transcriptase enzyme or kit according to the requirements of your RNA template or application.
|RNase H (+/-)
|Plus||Labeling for microarrays; specially designed for reverse transcription
with any amount of RNA; optimized buffer; no RNase H step needed
For full-length cDNAs up to 7 kb; starting material 10 pg–5 µg total RNA
Reverse Transcription Kit
|Minus|| Kit includes reverse transcriptase (RT32) and RNase inhibitor, 2x Master Mix
and RNase H (for optional step)
RNA polymerases and ligases
RNA polymerases, or more specifically DNA-directed RNA polymerases, are enzymes that synthesize RNA from a DNA template. The enzyme Poly(A) polymerase uses single-stranded RNA as a primer to add a poly(A) tail to RNA by catalyzing the incorporation of adenine residues into the 3’ termini of RNA.
T4 RNA ligases are useful enzymes for RNA analysis particularly upstream of procedures such as high-throughput RNA sequencing and microarrays. T4 RNA ligases 1 and 2 are enzymes that can label, circularize or perform intermolecular ligation of RNA by joining adjacent 3'-OH and 5'-PO4 polynucleotides. Attachment of adapters to RNA 3'-ends with T4 RNA ligase 1 is a useful first step for RNA quantification and discovery by RT-PCR and high-throughput sequencing.
Manipulating RNA is made easy with these polymerases and ligases.
Digesting RNA and protecting RNA
Ribonuclease protection assay (RPA) with the enzyme RNase A is a technique used to determine relative or absolute transcript abundance and to map mRNA termini and intron/exon boundaries.
Single base substitutions can be detected and localized by a simple enzyme method that involves RNase A cleavage of single base mismatches in RNA:DNA heteroduplexes.
|Endoribonuclease that degrades ssRNA at C and
RNase H (BLIRT)
|Endoribonuclease that specifically hydrolyzes
the phosphodiester bonds of RNA hybridized
RNase Inhibitor Hu
|Human placental protein inhibits ribonuclease
(RNase) activity of common eukaryotic enzymes
such as RNase A, RNase B, RNase C
Discover featured enzymes for RNA analysis
FAQs about RNA analysis
Starting a new lab? Your nearest sales representative is waiting to hear from you.