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11277073910
Roche
DIG RNA Labeling Mix
sufficient for 20 reactions, solution
Synonim(y):
znakowanie kwasów nukleinowych
Zaloguj sięWyświetlanie cen organizacyjnych i kontraktowych
About This Item
Kod UNSPSC:
41105500
Polecane produkty
Formularz
solution
Poziom jakości
zastosowanie
sufficient for 20 reactions
opakowanie
pkg of 40 μL
producent / nazwa handlowa
Roche
zanieczyszczenia
Ribonuclease, none detected (up to 20 µl using MSII-RNA)
kolor
colorless
rozpuszczalność
water: miscible
temp. przechowywania
−20°C
Opis ogólny
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.
Specyficzność
Heat inactivation: Stop the reaction by adding 2 μl 0.2 M EDTA (pH 8.0).
Zastosowanie
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
Cechy i korzyści
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.
Jakość
Function tested in the DIG RNA Labeling Kit and in the DIG Nucleic Acid Detection Kit.
Inne uwagi
For life science research only. Not for use in diagnostic procedures.
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Hasło ostrzegawcze
Warning
Zwroty wskazujące rodzaj zagrożenia
Zwroty wskazujące środki ostrożności
Klasyfikacja zagrożeń
Acute Tox. 4 Oral
Kod klasy składowania
12 - Non Combustible Liquids
Klasa zagrożenia wodnego (WGK)
WGK 1
Temperatura zapłonu (°F)
does not flash
Temperatura zapłonu (°C)
does not flash
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Produkty
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.
Metody znakowania digoksygeniną (DIG) i zestawy do sond DNA i RNA DIG, znakowanie DNA z losowym primerem, znakowanie nickiem translacyjnym, znakowanie końcowe oligonukleotydów 5' i 3'.
Protokoły
DIG RNA Labeling Mix Protocol & Troubleshooting
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