RNA-seq Analysis of TERC Transcripts
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Corresponding Organization :
Other organizations : Boston Children's Hospital, Dana-Farber Cancer Institute, Tufts University
Protocol cited in 5 other protocols
Variable analysis
- Amount of total RNA (600 ng)
- Concentration of 5'-adenylated, 3'-blocked adaptor (5 μM)
- Amount of T4 RNA ligase, Truncated KQ (280 units)
- Concentration of PEG 8000 (25%)
- Amount of RNaseOUT (1 μl)
- Reaction time (16-24 h)
- Concentration of universal RT primer (5 pmol)
- Concentration of TERC_L2 and universal RT or TERC_L3 and universal RT primer sets (5 μM)
- Ligation of total RNA to the adaptor
- Synthesis of cDNA
- Amplification of mature TERC and extended TERC transcripts
- Visualization of PCR products on agarose gels
- Sequencing of 3' RACE PCR products
- Temperature (25 °C)
- Enzyme used for cDNA synthesis (SuperScript III reverse transcriptase)
- None specified
- None specified
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