추천 제품
양식
solution
Quality Level
사용
sufficient for 20 reactions
포장
pkg of 40 μL
제조업체/상표
Roche
불순물
Ribonuclease, none detected (up to 20 µl using MSII-RNA)
색상
colorless
solubility
water: miscible
저장 온도
−20°C
일반 설명
Labeling efficiency: Approximately 10μg of full-length digoxigenin-labeled RNA is transcribed from 1μg linear template DNA.
Assay Time: 135 minutes
Sample Materials
Linearized plasmid DNA:
The DNA to be transcribed is cloned into the polylinker site of an appropriate transcription vector which contains adjacent to the polylinker a promoter for SP6, T7 or T3 RNA polymerase. For the synthesis of ‘run off′ transcripts the plasmid is linearized by a restriction enzyme. Restriction enzymes creating 5′-overhangs should be used; 3′ overhangs should be avoided. The linearized template DNA should be purified by phenol chloroform extraction and ethanol precipitation, to avoid RNase contamination. For ′run around′ transcription circular plasmid DNA is used.
PCR product:
PCR-fragments which contain RNA polymerase promoter sequences can also be used as templates for transcription. Purification of the PCR fragment by HighPure column purification prior to transcription is recommended.
Assay Time: 135 minutes
Sample Materials
Linearized plasmid DNA:
The DNA to be transcribed is cloned into the polylinker site of an appropriate transcription vector which contains adjacent to the polylinker a promoter for SP6, T7 or T3 RNA polymerase. For the synthesis of ‘run off′ transcripts the plasmid is linearized by a restriction enzyme. Restriction enzymes creating 5′-overhangs should be used; 3′ overhangs should be avoided. The linearized template DNA should be purified by phenol chloroform extraction and ethanol precipitation, to avoid RNase contamination. For ′run around′ transcription circular plasmid DNA is used.
PCR product:
PCR-fragments which contain RNA polymerase promoter sequences can also be used as templates for transcription. Purification of the PCR fragment by HighPure column purification prior to transcription is recommended.
DIG-labeled, single-stranded RNA probes of defined length are generated by in vitro transcription. DIG-11-UTP is incorporated by SP6, T7, and T3 RNA polymerases at approximately every 20 to 25th nucleotide of the transcript under standard conditions. The DIG RNA Labeling Mix is specifically designed for the use with SP6, T7, and T3 RNA polymerases, which are supplied with an optimized transcription buffer.
Convenient nucleotide mixture for the labeling of RNA with Digoxigenin-11-UTP.
Contents
10x solution with: 10 mM ATP, CTP, GTP (each), 6.5 mM UTP, 3.5 mM DIG-11-UTP.
Convenient nucleotide mixture for the labeling of RNA with Digoxigenin-11-UTP.
Contents
10x solution with: 10 mM ATP, CTP, GTP (each), 6.5 mM UTP, 3.5 mM DIG-11-UTP.
특이성
Heat inactivation: Stop the reaction by adding 2 μl 0.2 M EDTA (pH 8.0).
애플리케이션
RNA labeling with Digoxigenin-11-UTP by in vitro transcription with SP6, T7, and T3 RNA polymerases. DIG-labeled RNA is used in a variety of hybridization techniques:
- Northern blots
- Southern blots
- Dot blots
- Plaque or colony lifts
- RNase protection experiments
- Chromosomes, cells, and tissue sections in situ
특징 및 장점
The DIG RNA Labeling Mix is especially designed for the use with SP6,T7 and T3 RNA polymerases from Roche which are supplied with an optimized transcription buffer.
품질
Function tested in the DIG RNA Labeling Kit and in the DIG Nucleic Acid Detection Kit.
기타 정보
For life science research only. Not for use in diagnostic procedures.
신호어
Warning
유해 및 위험 성명서
Hazard Classifications
Acute Tox. 4 Oral
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 1
Flash Point (°F)
does not flash
Flash Point (°C)
does not flash
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문서
Digoxigenin (DIG) labeling methods and kits for DNA and RNA DIG probes, random primed DNA labeling, nick translation labeling, 5’ and 3’ oligonucleotide end-labeling.
프로토콜
Determine the labeling efficiency in terms of μg (expected yield of a standard labeling reaction is 20 μg of DIG labeled RNA per μg linearized template DNA after the DIG RNA labeling reaction).
자사의 과학자팀은 생명 과학, 재료 과학, 화학 합성, 크로마토그래피, 분석 및 기타 많은 영역을 포함한 모든 과학 분야에 경험이 있습니다..
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