[PM1600] ExcelBand™ All Blue Regular Range Plus Protein Marker (9-180 kDa), 250 μl x 2
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The PM1600 ExcelBand™ All Blue Regular Range Plus Protein Marker is a blue protein standard with 11 pre-stained proteins covering a wide range of molecular weights from 10 to 180 kDa in Tris-Glycine buffer (9 to 170 kDa in Bis-Tris (MOPS) buffer and Bis-Tris (MES) buffer). Proteins are covalently coupled with a blue chromophore, and two reference bands (at 25 kDa and 72 kDa, respectively) are enhanced in intensity when separated on SDS-PAGE (Tris-Glycine buffer).
The PM1600 ExcelBand™ All Blue Regular Range Plus Protein Marker is designed for monitoring protein separation during SDS-polyacrylamide gel electrophoresis, verification of Western transfer efficiency on membranes (PVDF, nylon, or nitrocellulose) and for approximating the size of proteins.
Detail
Description
The PM1600 ExcelBand™ All Blue Regular Range Plus Protein Marker is a blue protein standard with 11 pre-stained proteins covering a wide range of molecular weights from 10 to 180 kDa in Tris-Glycine buffer (9 to 170 kDa in Bis-Tris (MOPS) buffer and Bis-Tris (MES) buffer). Proteins are covalently coupled with a blue chromophore, and two reference bands (at 25 kDa and 72 kDa, respectively) are enhanced in intensity when separated on SDS-PAGE (Tris-Glycine buffer).
The PM1600 ExcelBand™ All Blue Regular Range Plus Protein Marker is designed for monitoring protein separation during SDS-polyacrylamide gel electrophoresis, verification of Western transfer efficiency on membranes (PVDF, nylon, or nitrocellulose) and for approximating the size of proteins.
Features
Ready-to-use — Premixed with a loading buffer for direct loading, no need to boil.
Two enhanced bands — 72 kDa and 25 kDa
Contents
Approximately 0.1~0.5 mg/ml of each protein in the buffer (20 mM Tris-phosphate (pH 7.5 at 25°C), 2% SDS, 0.2 mM DTT, 3.6 M urea, and 15% (v/v) glycerol).
Quality Control
Under suggested conditions, PM1600 ExcelBand™ All Blue Regular Range Plus Protein Marker resolves 11 major bands in 15% SDS-PAGE (Tris-Glycine buffer, MOPS, and MES buffer) and after Western blotting to nitrocellulose membrane.
Excellent separation and purification of cytoplasmic and nuclear RNA
Convenient and fast spin column format
High quality and yield of RNA
Isolate full diversity of RNA (including microRNA) without phenol
Purified RNA is ready for any application including RT-PCR, qRT-PCR, RNA-Seq, arrays and more
Cytoplasmic RNA is free of DNA and ready for direct use in RT-PCR/qRT-PCR
Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix
This kit provides a rapid method for the isolation and purification of both cytoplasmic and nuclear RNA from cultured animal cells and small tissue samples. The kit can be used to isolate all sizes of RNA from the cytoplasmic and nuclear RNA fractions, including all small RNA species without any requirement for phenol. Included in the kit are sufficient reagents to perform either 50 cytoplasmic RNA preparations or 25 cytoplasmic and 25 nuclear RNA preparations. Ten samples can be processed in approximately 45 minutes. This kit is also available in a 100 prep size.
Background
In certain circumstances it is desirable to be able to isolate fractionated RNA as opposed to total RNA. For example, it may be preferable to isolate only mature, cytoplasmic RNA for some studies on expression profiling. Alternatively it may be desirable to isolate nuclear RNA in order to investigate and study pre-processed (non-spliced) RNA. Furthermore, this kit can be used to isolate RNA for downstream applications where it is necessary to avoid DNA contamination, since the cytoplasmic fraction has been shown to be free of all traces of genomic DNA.
RNA Yield HeLa (1 x 106) – Cytoplasmic RNA HeLa (1 x 106) – Nuclear RNA
15 µg ≤ 3.5 µg
Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
Cod Uracil-DNA Glycosylase (Cod UNG) from Atlantic Cod is the only commercially available UNG enzyme that is completely and irreversibly inactivated by moderate heat treatment. The enzyme is produced in a recombinant E. coli (ung-) strain that contains a modified Cod UNG gene.
Key Features:
Heat-labile – Completely and irreversibly inactivated at 55°C
Contamination control – ideal in applications below
Use of Cod UNG makes contamination control possible in RT-PCR
Does not degrade PCR product post-PCR. This makes downstream use of the PCR product possible
High purity enzyme, tested free of contaminating nucleases
There are several commercially available Uracil-DNA glycosylases on the market today. Most of them are of bacterial origin and work well if you have no intention to further analyze the PCR products post-PCR. However, if you want to store your PCR products for downstream analysis such as cloning and sequencing, the reactivation of UNG and subsequent degradation of your PCR products are a problem with most of the commercially available UNGs. Cod UNG from ArcticZymes is completely and irreversibly inactivated by heat thus ensuring that sample integrity is maintained long-term regardless of storage conditions.
This is illustrated in figure 1, below
Figures
Properties
Recommended Protocols
1. Contamination control in PCR, qPCR and one-step RT-qPCR
Cod UNG works in all commercially available master mixes.
Be sure that you have used dUTP containing dNTP mixes in your previous PCR experiments.
Add 0.2 U Cod UNG directly to your 20 µl PCR reaction.
pre-incubate for 5 min at room temperature.
For RT-qPCR, reverse transcribe your RNA at 50-55°C.
Run your PCR.
Store your PCR product at -20°C or 4°C degrees.
2. Contamination control in RT-LAMP
Cod UNG is ideal for contamination control in RT-LAMP. One unit of Cod UNG per 30 μl reaction is sufficient for removing even high concentrations of carry-over contamination.
Ensure that you use dNTP mixes containing dUTP in your experiments.
Check that the RT-LAMP reaction is compatible with dUTP by running side-by-side reactions containing different ratios of dUTP to dUTP (100% dUTP, 90% dUTP, 80% dUTP and 0% dUTP).
Add 1 U Cod UNG directly to your 30 µl RT-LAMP reaction.
Prepare the reaction mix on ice.
Analyze your RNA at 65°C, no preincubation is necessary.
Cod Uracil-DNA Glycosylase (Cod UNG) from Atlantic Cod is the only commercially available UNG enzyme that is completely and irreversibly inactivated by moderate heat treatment. The enzyme is produced in a recombinant E. coli (ung-) strain that contains a modified Cod UNG gene.
