The UltraMarathonRTTM Template Switching Kit is ideal for library prep for RNA-seq applications. It has consistent and efficient terminal transferase activity, allowing it to be used in applications where template switching is preferred. uMRT can create cDNA from any RNA: long or short RNAs, highly structured or difficult transcripts of any abundance. uMRT used with our Template Switching Oligo (TSO) will allow you to create full-length cDNAs while adding appropriate sequencing adapters. Use the uMRT Template Switching Kit to reduce cDNA library bias.
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:
- UltraMarathonRT
- 2x RT Reaction Buffer
- 5x Template Switching Buffer
- uMRT-TSO
- Low Boost
- Nuclease-free water
Catalog# R1004S (20 reactions), R1004M (50 reactions), R1004L (100 reactions)
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- 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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