Rapid Amplification of cDNA Ends (RACE)는 RNA의 full-length sequence를 얻기 위해 사용되는 기술이다. RACE는 RNA의 일부 알고 있는 부위 (small known sequence)를 이용하여 미지의 5’ 말단 (5’ RACE-PCR법), 3’ 말단 (3’ RACE-PCR법)을 규명하는데 적합한 기술이다.
본 제품 SMARTer RACE 5’/3’ Kit은 SMARTer 기술을 이용한 RACE cDNA 합성 키트로써 별도의 adaptor ligation 없이도 짧은 시간 내에 full-length의 cDNA를 합성할 수 있다. 합성된 cDNA는 5’ RACE-PCR과 3’ RACE-PCR에 적용할 수 있다.
SMARTer RACE 5’/3’ Kit은 기존 SMARTer RACE cDNA Amplification Kit보다 더 낮은 background와 높은 특이성(higher specificity)을 보이는 제품이다. 또한 기존 제품에 비하여 농도가 낮은 RNA 샘플 (Input volume up to 11ul)도 사용 가능하며, Long fragment이거나 GC-rich template와 같은 증폭이 어려운 서열도 효율적으로 증폭 가능하다. 본 제품 내에는 SeqAmp DNA Polymerase, In-Fusion HD Cloning kit, linearized vector, competent cell까지 포함되어 있다.
특징
- Easy to Use - performed in a single tube, and minimizes handling of both your RNA sample and your synthesized cDNA.
- Complete Kit - contains all the necessary components (save gene-specific primers) to streamline your process.
- Requires Only 10 ng of Total RNA - utilize small samples, including biopsies, tissue dissections, needle aspirations,
and embryonic and rare disease tissues.
- Specific Enrichment for 5' Ends
Our carefully designed, chemically-treated SMARTer Oligo preferentially hybridizes to the 5' ends of the cDNA being synthesized. Using
this SMARTer Oligo, our procedure enriches cDNA pools for 5' sequences.
- No RNA Pretreatment Required - requires no RNA pretreatment with DNase. This protocol works with total RNA, as well
as with samples that may be contaminated with genomic DNA.
그림 1. Overview of the SMARTer RACE 5'/3' Kit workflow. Each kit is a complete system, containing the reagents
required to recover cloned RACE fragments on the second day.
그림 2. SMART(er) cDNA synthesis compared to conventional cDNA synthesis.
그림 3. SMARTer 5'- and 3'-RACE results for different genes. Lane 1: 1 kb DNA marker. Lane 2: 100 bp DNA marker.
Lanes 3 & 4: tyrosyl-tRNA synthetase (YARS). Lanes 5 & 6: interferon gamma receptor 1 (IFNGR1). Lanes 7 & 8: adrenergic,
beta, receptor kinase 1 (ADRBK1). Lanes 9 & 10: actin, beta (ACTB). Lanes 11 & 12: transferrin receptor
(TFR) Lane 13: low density lipoprotein-related protein 2 (LRP2). Lane 14: low density lipoprotein-related protein 1 (LRP1).
적용
- RACE (Rapid Amplification of cDNA Ends)
- Identification of 3'/5' UTRs
- Full-length gene cloning
- Gene cloning and analysis
구성품 (자세한 내용은 CoA를 참조하세요)
* SMARTer RACE 5’/3’ Kit Components (Cat. No. 634860) (Not sold separately)
* SeqAmp DNA Polymerase (Cat. No. 638504)
* In-Fusion HD Cloning Kit (Cat. No. 639648) (Not sold separately)
* NucleoSpin Gel and PCR Clean-Up Kit (Cat. No. 740609.10) (Not sold separately)
* Stellar Competent Cells (Cat. No. 636763)