UltraMarathonRTTM Template Switching Kit can provide full length cDNAs even when you do not know the sequences at either end of the RNA transcript. Its terminal transferase activity allows for template switching when used with the uMRT Template Switching Oligonucleotide. It is designed to capture low abundance RNA with high efficiency and sensitivity and uMRT Boost allows uMRT to capture highly structured long RNAs at ultra-low input.
Used for bulk sequencing library preparation and low abundant RNA samples in long-read sequencing, SMART-seq, and RACE applications (use with RNA input 0.1 ng – 20 ng).
UltraMarathonRT Template Switching Kit Components:
- uMRT
- RT Buffer
- Template Switching Buffer
- uMRT Template Switching Oligonucleotide
- uMRT Boost
- Nuclease-free water
For purchase orders please contact us at sales@rnaconnect.com.
- dNTP mix stock (10 mM each)
- The dNTP mix contains premixed aqueous solutions of dATP, dCTP, dGTP and dTTP, each at a final concentration of 10 mM. High purity nucleotides (>99%) are recommended, and the pH should be adjusted to 7.0 with sodium hydroxide.
- dATP stock (10 mM)
- dATP is needed for the non-templated addition of three adenosines (AAA) to the 3'-end of cDNA primer extension products that facilitates the template switching process.
- Custom oligonucleotides
- uMRT dT18 primer
- Custom oligo-dT primer needed for reverse transcription.
- Custom primer used for full-length cDNA preamplification. (Custom primer sequences are provided in the template switching protocol)
- AmpPCR primer
- uMRT dT18 primer
- KAPA HiFi DNA polymerase (Roche, Cat# KK2102) is the recommended DNA polymerase as it provides the most full-length cDNA products
Storage conditions: For short duration of storage (< 3 month), UltraMarathonRT can be placed in a -20℃ freezer. -80℃ is recommended for long-term storage. Note that UltraMarathonRT can endure 10 freeze-thaw cycles without losing noticeable activity. After 20 freeze-thaw cycles, UltraMarathonRT maintains 90% of its original activity.
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