• Second Strand cDNA Synthesis Kit
  • Second Strand cDNA Synthesis Kit

Second Strand cDNA Synthesis Kit

This product is a reagent kit that removes RNA strands from RNA DNA heterozygous strands on the basis of synthesizing the first strand of cDNA and synthesizes the second strand of cDNA, ultimately forming a double stranded cDNA. This kit contains various reagents required for cDNA second strand synthesis.
$1.70
  • Second Strand cDNA Synthesis Kit

SPECIFICATION

            Second Strand cDNA Synthesis Kit
This product is a reagent kit that removes RNA strands from RNA DNA heterozygous strands on the basis of synthesizing the first strand of cDNA and synthesizes the second strand of cDNA, ultimately forming a double stranded cDNA. This kit contains various reagents required for cDNA second strand synthesis.
This kit uses RNase H to induce cleavage of RNA in RNA DNA heterozygous strands. At this time, DNA Polymerase I can catalyze the formation of cDNA second strands through cleavage translation reaction. The double stranded cDNA synthesized using this kit can be directly used for subsequent routine PCR, real-time PCR, cDNA library construction, etc.
Component specifications
RNase H (1U/ μ L) 10 μ L
DNA Polymerase I (10U/ μ L) 30 μ L
ten × Action Buffer 100 μ L
Storage: -20 ℃
When conducting the second chain synthesis, the appropriate final concentration of dNTP is about 0.2mM. If the final concentration is too low, it is necessary to supplement dNTP appropriately in the reaction system.
If you want to obtain flat ended double stranded cDNA, you can use T4 DNA Polymerase (YT403) or DNA End Smoothing Kit (YT404) for processing.
Enzymes should be stored in an ice box or ice bath when used, and should be immediately stored at -20 ℃ after use.
This product is limited to scientific research by professionals and cannot be used for clinical diagnosis or treatment, nor for food or medicine.
For your safety and health, please wear laboratory clothes and disposable gloves for operation.
Synthesize the first strand of cDNA using reverse transcriptase, such as BalbRT II M-MLV reverse transcriptase (YT394) or BalbRT II cDNA first strand synthesis kit (YT397), and incubate at 70 ℃ for 10 minutes to terminate the reaction.
20 that have already synthesized the first chain on an ice bath μ Add the following reagents in sequence to the L reaction system:
Ingredient dosage
ten × Action Buffer 8 μ L
Nuclease free deionized water 68 μ L
RNase H (1U/ μ L) 1 μ L
DNA Polymerase I (10U/ μ L) 3 μ L
After preparing according to the above system, gently mix (you can use a pipette to blow and mix or Vortex to gently mix at the lowest speed), and then centrifuge to precipitate the liquid.
Incubate at 15 ℃ for 2 hours.
Attention: The reaction temperature should not exceed 15 ℃
Add 5 μ Mix well with L 0.5M EDTA (pH8.0) to terminate the reaction.
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