ERT-RO

Expand^™ Reverse Transcriptase

solution, recombinant, expressed in E. coli (AP401 (k)), sufficient for 20 reactions, sufficient for 100 reactions

• Synonym(s): reverse transcriptase, expand
• Enzyme Commission number: 2.7.7.49 (BRENDA, IUBMB)

Properties

• recombinant: expressed in E. coli (AP401 (k))
• Quality Level: 100
• form: solution
• usage: sufficient for 100 reactions; sufficient for 20 reactions
• packaging: pkg of 1,000 U (11785826001 [20 μl]); pkg of 5,000 U (11785834001 [100 μl])
• manufacturer/tradename: Roche
• parameter: 42-43 °C optimum reaction temp.
• technique(s): RT-PCR: suitable; RT-qPCR: suitable
• input: purified RNA
• detection method: probe-based
• storage temp.: −20°C

Description

General description

Expand^™ Reverse Transcriptase is a RNA directed DNA polymerase optimized for amplification of cDNA fragments up to 13.5 kb in a two-step RT-PCR assay. The enzyme is a genetically engineered version of the Moloney Murine Leukemia Virus reverse transcriptase (M-MuLV-RT). A point mutation within the RNase H sequence reduces RNase H activity to undetectable levels. Detectable improvements are obtained with respect to higher amounts of full-length cDNA transcripts and longer transcripts compared to using the native M-MuLV-RT.

Specificity

Heat inactivation: 95 °C for 2 min
Reverse Transcriptase enzyme for RT-PCR reactions up to 3 kb

Application

Expand^™ Reverse Transcriptase has been used in real-time PCR analysis for conversion of isolated RNA into cDNA. It has also been used in reverse transcription (RT). Expand^™ Reverse Transcriptase may also be used for analyzing the sequence of the cDNA and comparing it to the genomic sequence. Important information can be obtained on intervening sequences (introns) and genomic recombinations.

Features and Benefits

• Obtain higher amounts of full-length cDNA transcripts and longer transcripts up to 13.5kb
• No RNase H activity
• Efficiently transcribes total RNA, mRNA, viral RNA and RNA rich in secondary structures
• Ideal for two-Step RT-PCR assay
• Purification of cDNA before PCR reaction is not necessary

Packaging

1 set containing 3 components

Unit Definition

One unit is the enzyme activity that incorporates 1.0 nmol TMP into acid insoluble products in 10 min at +37 °C with poly(A)⁺ × (dT)¹⁵ as substrate.

Volume Activity: 50 U/μl

Other Notes

For life science research only. Not for use in diagnostic procedures.

Legal Information

Expand is a trademark of Roche

Kit Components Only

• Expand Reverse Transcriptase 50 U/μl
• Expand Reverse Transcriptase Buffer 5x concentrated
• DTT Stock Solution 100 mM

Safety Information

• Pictograms: GHS07
• Signal Word: Warning
• Hazard Statements: H319
• Precautionary Statements: P264 - P280 - P305 + P351 + P338 - P337 + P313
• Hazard Classifications: Eye Irrit. 2
• Storage Class Code: 12 - Non Combustible Liquids
• WGK: WGK 1
• Flash Point(C): does not flash