1. SweScript RT I First Strand cDNA Synthesis Kit, 50 rxns $399,

Product Information

Description/Introduction

This product utilizes the modified mutant reverse transcriptase to synthesize the first strand of cDNA with total RNA or mRNA as the template for efficient reverse transcription. The kit contains all the reagents needed to synthesize the first strand of cDNA with high quality, and provides two kinds of cDNA synthesis primers for choice: The Random Hexamer Primer and Oligo (dT)18 Primer. The synthesized single-stranded cDNA products can be directly used for subsequent PCR or qPCR. SWEScript RT I (Reverse Transcriptase) is a mutant Reverse Transcriptase based on M-MLV Reverse Transcriptase and obtained through in vitro evolutionary screening. SWEScript RT I has no RNase H activity. So the RNA degradation in the template of DNA/RNA hybridization during the first strand cDNA synthesis reaction can be avoided, so as to ensure the amount and length of the first strand cDNA synthesis. Compared with the wild-type reverse transcriptase, SweScript RT I has improved thermal stability and cDNA synthesis efficiency. It can efficiently synthesize the first strand cDNA at the range of 42-55℃, and the length of cDNA can acheive to 10 kb.

Storage and Handling Conditions

Wet ice bag transportation; Stored at -20℃ temperature, valid for 12 months.

Component

a：with RNase Inhibitor

b：with dNTP Mixture & Mg²+

Assay Protocol

1. Preparation of reverse transcription reaction system (20ul reaction system is recommended);

Note: For high GC or complex templates, it is recommended that the RNA template, reverse transcription primers and nucrelease-free water can be mixed in advance and then cooled on ice quickly after being kept at 65℃for 5 min. And then you add the other components.

2. Gently mix and centrifuge;

3. Reverse transcription program setting:

a: For example, the Random Hexamer Primer is selected and incubated at 25℃for 5 min, then the subsequent reaction is carried out. If you choose to use Oligo (dT)18 Primer or Gene Specific Primer, you can directly react at 50℃.

b: For high GC or complex templates, the reverse transcription temperature can be raised to 55℃.

Note:

1. Please wear disposable gloves during operation to avoid RNase contamination.

2. The reverse transcription products can be stored at -20℃ for a period. If long-term storage is needed, it is recommended to store them at -80℃ after packing and avoid repeated freeze-thaw cycles.

3. If the template is of eukaryotic origin, it is recommended to select Oligo (dT)18 Primer and pair it with the 3′ Poly A tail of eukaryotic mRNA to obtain the highest yield of full-length cDNA.

4. For reverse transcription of prokaryotic RNA, Random Hexamer Primer or Gene Specific Primer should be used.

5. Random Hexamer Primer has wide applicability and is suitable for mRNA, rRNA, tRNA, small RNA and lncRNA templates.

6. If reverse transcription is followed by qPCR assay, Oligo (dT)18 Primer and Random Hexamer Primer can be mixed to make cDNA synthesis efficiency in all regions of mRNA the same, which helps to improve the authenticity and repeatability of quantitative results.

For Research Use Only!