추천 제품
생물학적 소스
Escherichia coli (BL 21/pAR 1219)
Quality Level
분석
100% (SDS-PAGE)
형태
solution
특이 활성도
≥20 U/μL
분자량
98 kDa (Single polypeptide chain)
포장
pkg of 1,000 U (10881767001)
pkg of 5,000 U (10881775001)
제조업체/상표
Roche
기술
Northern blotting: suitable
Southern blotting: suitable
hybridization: suitable
색상
colorless
pH
7.9 (39 °F)
solubility
water: miscible
적합성
suitable for molecular biology
NCBI 수납 번호
UniProt 수납 번호
응용 분야
life science and biopharma
외래 활성
Endonucleases 100 units, none detected
Nicking activity 100 units, none detected
RNase 100 units, none detected
저장 온도
−20°C
유전자 정보
Escherichia coli ... T7p07(1261050)
일반 설명
T7 RNA polymerase is commonly used to transcribe DNA which has been cloned into vectors which have two phage promoters in opposite orientation. RNA can be selectively synthesized from either strand of the insert DNA with different polymerases. Homogeneously labeled single-stranded RNA can be generated with this system. Transcripts can be non-radioactively labeled with biotin or DIG-11-UTP or radioactively labeled to high specific activity with [α-32P] or [α-35S]-labeled nucleotides.
Synthesis of hybridization probes: T7 RNA polymerase allows highly efficient production of homogeneously labeled RNA. This labeled RNA may be used as hybridization probes in Southern, northern, and dot blots, as well as in situ hybridizations.
Suitable labels:Transcripts can be nonradioactively labeled with biotin-16-UTP or DIG-11-UTP. They may also be radioactively labeled to high specific activity with [α-32P]- or [α-35S]-labeled nucleotides.
Synthesis of hybridization probes: T7 RNA polymerase allows highly efficient production of homogeneously labeled RNA. This labeled RNA may be used as hybridization probes in Southern, northern, and dot blots, as well as in situ hybridizations.
Suitable labels:Transcripts can be nonradioactively labeled with biotin-16-UTP or DIG-11-UTP. They may also be radioactively labeled to high specific activity with [α-32P]- or [α-35S]-labeled nucleotides.
특이성
Promotor specifity
T7 RNA polymerase is extremely promoter-specific and only transcribes bacteriophage T7 DNA or DNA cloned downstream of a T7 promoter. Although the T7 and T3 promoter sequences differ only by 3 bp, T7 RNA polymerase only transcribes DNA cloned downstream of its promoter.
Heat inactivation: Stop the reaction by adding 2 μl 0.2 M EDTA (pH 8.0) and/or heat to 65 °C.
T7 RNA polymerase is extremely promoter-specific and only transcribes bacteriophage T7 DNA or DNA cloned downstream of a T7 promoter. Although the T7 and T3 promoter sequences differ only by 3 bp, T7 RNA polymerase only transcribes DNA cloned downstream of its promoter.
Heat inactivation: Stop the reaction by adding 2 μl 0.2 M EDTA (pH 8.0) and/or heat to 65 °C.
애플리케이션
- T7 RNA Polymerase can transcribe RNA from cloned DNA templates that are downstream from a T7 promoter. The synthesis can be performed with labeled NTPs to generate highly labeled RNA. Synthesized RNA can be used in many applications, including: RNA or DNA blotting techniques
- In situ hybridization
- RNase protection studies: Transcripts synthesized by the enzyme are used as precursor RNA for studies on RNA splicing and processing.
- Synthesis of capped RNA in vitro with addition of m7GpppG or m7GpppA in excess over GTP or ATP during the transcription reaction. The generated antisense RNA can be introduced into cells to suppress the expression of the corresponding genes.
- Microarray target synthesis
포장
1 kit containing 2 components
단위 정의
One unit is the enzyme activity which incorporates 1 nmol CMP in acid-precipitable RNA products within 60 minutes at +37 °C.
Volume Activity: ≥20 U/μl
Volume Activity: ≥20 U/μl
제조 메모
Activator: The T7 RNA polymerases are strongly stimulated by BSA or spermidine.
저장 및 안정성
Keep container tightly closed in a dry and well-ventilated place.
기타 정보
Test Buffer
40 mM Tris-HCl, pH 8.0 (+20°C), 6 mM MgCl2, 10 mM dithiothreitol, 2 mM spermidine, pH approximately 8.0 (+20°C).
Absence of Endonucleases
1. 1 μg lambda DNA is incubated with T7 RNA polymerase for 4 hours at +37°C in 25 μl test buffer. The number of enzyme units which show no degradation of lambda DNA is >100 U.
2. 1 μg Eco RI/Hind III fragments of lambda DNA is incubated with T7 RNA polymerase for 4 hours at +37°C in 25 μl test buffer. The number of enzyme units which show no alteration of the banding pattern is >100 U.
Absence of Nicking Activity
1 μg pBR322 DNA is incubated with T7 RNA polymerase for 4 hours at +37°C in 25 μl test buffer. The number of enzyme units which show no relaxing of supercoiled structure is >100 U.
Absence of RNases
4 μg MS2 RNA are incubated with T7 RNA polymerase for 4 hours at +37°C in 50 μl test buffer. The number of enzyme units which show no degradation of MS2 RNA is >100 U.
Performance in Transcription Assay
T7 RNA polymerase is function tested in the SP6/T7 Transcription Kit (Cat. No. 10 999 644 001). The incorporation rate in the standard assay with 0.5 μg pSPT19 neo DNA linearized with Eco RI and 50 mCi [alpha-32P] CTP, [400 Ci/mmol (15 TBq/mmol)] gives >50% of the input radioactivity in 20 minutes.
40 mM Tris-HCl, pH 8.0 (+20°C), 6 mM MgCl2, 10 mM dithiothreitol, 2 mM spermidine, pH approximately 8.0 (+20°C).
Absence of Endonucleases
1. 1 μg lambda DNA is incubated with T7 RNA polymerase for 4 hours at +37°C in 25 μl test buffer. The number of enzyme units which show no degradation of lambda DNA is >100 U.
2. 1 μg Eco RI/Hind III fragments of lambda DNA is incubated with T7 RNA polymerase for 4 hours at +37°C in 25 μl test buffer. The number of enzyme units which show no alteration of the banding pattern is >100 U.
Absence of Nicking Activity
1 μg pBR322 DNA is incubated with T7 RNA polymerase for 4 hours at +37°C in 25 μl test buffer. The number of enzyme units which show no relaxing of supercoiled structure is >100 U.
Absence of RNases
4 μg MS2 RNA are incubated with T7 RNA polymerase for 4 hours at +37°C in 50 μl test buffer. The number of enzyme units which show no degradation of MS2 RNA is >100 U.
Performance in Transcription Assay
T7 RNA polymerase is function tested in the SP6/T7 Transcription Kit (Cat. No. 10 999 644 001). The incorporation rate in the standard assay with 0.5 μg pSPT19 neo DNA linearized with Eco RI and 50 mCi [alpha-32P] CTP, [400 Ci/mmol (15 TBq/mmol)] gives >50% of the input radioactivity in 20 minutes.
For life science research only. Not for use in diagnostic procedures.
Roche has 10x concentrated RNA Labeling Mixes that are specially designed for DIG- or biotin-labeling. These mixes work well with T7 RNA Polymerase.
Roche has 10x concentrated RNA Labeling Mixes that are specially designed for DIG- or biotin-labeling. These mixes work well with T7 RNA Polymerase.
키트 구성품 전용
제품 번호
설명
- T7 RNA Polymerase, in buffer, pH 7.9 ≥20 U/μl
- Transcription Buffer 10x concentrated
신호어
Warning
유해 및 위험 성명서
Hazard Classifications
Eye Irrit. 2
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 2
Flash Point (°F)
does not flash
Flash Point (°C)
does not flash
시험 성적서(COA)
제품의 로트/배치 번호를 입력하여 시험 성적서(COA)을 검색하십시오. 로트 및 배치 번호는 제품 라벨에 있는 ‘로트’ 또는 ‘배치’라는 용어 뒤에서 찾을 수 있습니다.
이미 열람한 고객
Aditi Kulkarni et al.
Journal of genomics, 8, 30-36 (2020-03-20)
In the CRISPR-Cas systems, Cas13a is an RNA-guided RNA nuclease specifically targeting single strand RNA. We developed a Cas13a mediated CRISPR interference tool to target mRNA for gene silencing in mosquitoes. A Cas13a expressing plasmid was delivered to mosquitoes by
Kaia Achim et al.
Nature biotechnology, 33(5), 503-509 (2015-04-14)
Understanding cell type identity in a multicellular organism requires the integration of gene expression profiles from individual cells with their spatial location in a particular tissue. Current technologies allow whole-transcriptome sequencing of spatially identified cells but lack the throughput needed
Aditi Kulkarni et al.
Scientific reports, 12(1), 6005-6005 (2022-04-11)
Immune responses require delicate controls to maintain homeostasis while executing effective defense. Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor. The Krüppel-like factor 10 (KLF10) is a C2H2 zinc-finger containing transcription factor. The functions of mosquito AhR and KLF10
Min Li et al.
PloS one, 7(11), e49841-e49841 (2012-11-28)
IL-2 plays a key role in the survival and proliferation of immune cells, especially T lymphocytes. Its expression is precisely regulated at transcriptional and posttranscriptional level. IL-2 is known to be regulated by RNA binding proteins, such as tristetraprolin (TTP)
Camilo Riquelme-Guzmán et al.
eLife, 11 (2022-10-12)
Early events during axolotl limb regeneration include an immune response and the formation of a wound epithelium. These events are linked to a clearance of damaged tissue prior to blastema formation and regeneration of the missing structures. Here, we report
문서
T7 RNA Polymerase synthesizes RNA from DNA with high specificity, valuable for in vitro translation and anti-sense RNA.
Understand how mRNA vaccines induce immunity. and read how synthetic mRNA is prepared for vaccine immunogens and other biopharmaceuticals. Find reagents for synthesis of mRNA.
자사의 과학자팀은 생명 과학, 재료 과학, 화학 합성, 크로마토그래피, 분석 및 기타 많은 영역을 포함한 모든 과학 분야에 경험이 있습니다..
고객지원팀으로 연락바랍니다.