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Template Switching RT Enzyme Mix
The Template Switching RT Enzyme Mix and accompanying reaction buffer enable efficient template switching activity in a reverse transcription reaction. The mix contains a unique RT and Murine RNase Inhibitor. Unlike competitor RT products, no additives (such as PEG or betaine) are required for optimal performance, simplifying reaction setup. In conjunction with a template switching oligo (TSO), cDNA is synthesized with a known sequence of choice attached to the 3′ end. The resulting cDNA can be amplified by PCR or serve as template for 5′ RACE (rapid amplification of cDNA ends) or 2nd strand cDNA synthesis.
Prepare RNA-seq libraries from extremely low input: single cells/nuclei or 2 pg of total RNA.
Perform 5′ RACE from as low as 10 ng total RNA.
Low background for RNA-seq or 5′ RACE
Enzyme mix contains Murine RNase Inhibitor, no additives necessary
Compatible with various template switching oligos (TSOs), RT primers, and DNA polymerases for full length cDNA amplification
Enjoy faster protocols as compared to alternative RNA-seq methods (eg. Smart-Seq®)
产品信息
The reverse transcriptase (RT) in the Template Switching RT Enzyme Mix adds a few non-templated nucleotides after it reaches the 5′ end of the RNA template. These non-templated nucleotides can anneal to a template switching oligo (TSO) with a known sequence, prompting the reverse transcriptase to switch template from RNA to the TSO. The resulting cDNA contains a known sequence (complementary to the sequence of the TSO) attached to the 3′ end. This feature can be utilized in a variety of downstream applications, such as cDNA amplification, 5´ RACE (rapid amplification of cDNA ends), and 2nd strand cDNA synthesis. Optimized protocols are located in the Protocols section.
产品类别:
cDNA Synthesis & Reverse Transcriptases Products,
PCR, qPCR & Amplification Technologies Products
应用:
cDNA Synthesis,
RNA Analysis
产品组分信息
产品组分信息
本产品提供以下试剂或组分:
NEB #
名称
组分货号
储存温度
数量
浓度
M0466S
-20
Template Switching RT Enzyme Mix
M0466SVIAL
-20
1 x 0.02 ml
10 X
Template Switching RT Buffer
B0466SVIAL
-20
1 x 0.5 ml
4 X
M0466L
-20
Template Switching RT Enzyme Mix
M0466LVIAL
-20
1 x 0.1 ml
10 X
Template Switching RT Buffer
B0466SVIAL
-20
1 x 0.5 ml
4 X
特性和用法
操作说明、说明书 & 用法
操作说明
cDNA Synthesis and Amplification Protocol using the Template Switching RT Enzyme Mix (NEB #M0466)
2nd Strand cDNA Synthesis Protocol using the Template Switching RT Enzyme Mix (NEB #M0466)
5’ RACE Protocol using the Template Switching RT Enzyme Mix (NEB #M0466)
FAQs & 问题解决指南
FAQs
Do I need to add RNase Inhibitor to the RT reaction when using the Template Switching RT Enzyme Mix?
Numerous publications describe the addition of various additives (e.g., betaine and PEG) to improve template switching. Can I add any of those to my reaction?
Is the Template Switching RT Enzyme Mix compatible with RT primers with degenerate bases for cDNA synthesis and amplification, as described in the mcSCRB method [1]?
What type of template switching oligos (TSOs) are compatible with the Template Switching RT Enzyme Mix?
Can I use the Template Switching RT Enzyme Mix for regular RT reactions?
What is the efficiency of template switching when using the Template Switching RT Enzyme Mix?
Does the Template Switching RT Enzyme Mix work for prokaryotic RNA?
Does the Template Switching RT Enzyme Mix work for fragmented RNA?
What type of RT primers can be used with the Template Switching RT Enzyme Mix?