cDNA Synthesis for MicroRNA Profiling and Custom Gene Expression Taqman Arrays

The GRCF will prepare cDNA from customer supplied total RNA according to Life Technologies quick reference guide for Taqman OpenArray MicroRNA Panel Analysis or using a High Capacity cDNA Reverse Transcription Kit (Life Technologies part # 4368813) for the Gene Expression Array analysis on the QuantStudio 12K Flex Real-Time PCR System. The GRCF also has RNA isolation services available if you require start to finish processing.

cDNA synthesis using Superscript VILO (Life Technologies, catalog number 11754)

cDNA synthesis using a Superscript VILO kit is also available. We offer conversion of up to 2500 ng of RNA in one reaction. Greater than 2500 ng can be converted for an additional charge. The amount of RNA converted in one reaction is dependent on the concentration of the RNA.

RNA Requirements
  • RNA should be free of PCR inhibitors
  • Has a A260/230 ratio between 2.0 and 2.4
  • Has a A260/280 ratio between 1.8 and 2.1
  • Has a RNA Integrity Number (RIN) or RNA Quality Number (RQN) between 6.5 and 10

The recommended amount of total RNA for preparing cDNA for microRNA Profiling is 100ng.

The recommended amount of total RNA for preparing cDNA for Gene Expression Taqman Arrays is 10µl at 200ng/µl (2µg).

Qualitative analysis of your RNA samples is available by Advanced Analytical Fragment Analyzer CE analysis. The system determines a RQN (RNA Quality Number) of total RNA on a scale from 1 to 10 that correlates to current industry practices (RIN).

Pricing

$25-$27 per sample for cDNA synthesis

Ordering

Please contact Laura Kasch [410-614-3830] for additional information and ordering.

RT-PCR
RT-PCR is available using Qiagen’s PyroMark OneStep RT-PCR Kit or One-Step RT-PCR Kit

The Qiagen OneStep RT-PCR Kits provide highly sensitive and specific reverse transcription of any RNA template by using a blend of Sensiscript and Omniscript Reverse Transcriptases,, followed by highly specific and reliable amplification of cDNA using HotStarTaq DNA Polymerase. The entire process takes place in one tube reducing the risk of contamination. The RT-PCR product generated from the OneStep RT-PCR Kits can be used for Pyrosequencing applications such as genotyping and allele-specific gene expression analysis.

Pricing and Ordering

Pricing is project specific. Please contact Laura Kasch [410-614-3830] for additional information and ordering.