Introduction

This product provides fast and easy methods for purification of total DNA for reliable PCR and southern blotting. Total DNA can be purified from bacterial culture, body fluids, food and fermentation.

Details

Specifications

Features	Specifications
Main Functions	Isolation bacterial DNA from cultures, food and other samples
Applications	PCR, southern blot and enzyme digestion, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Culture medium, swab, parasitic blood, tissue,sputum, etc.
Sample amount	Bacterial culture medium: 0.5-2 mlTissue samples: 50-100 mg Whole blood / cell suspension: 0.5-1 ml Viscous secretion: 0.1-1 g
Elution volume	≥30μl
Time per run	≤40 minutes
Liquid carrying volume per column	800μl
Binding yield of column	100μg

Principle

This product is based on silica Column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption column. Nucleic acid is adsorbed on the membrane, while proteinis not adsorbed and is removed with filtration. After washing proteins and other impurities, Nucleic acid was finally eluted with low-salt buffer (10mmTris, pH9.0, 0.5mm EDTA). This product carries 0.1-0.2mm acid glass beads, which can improve the wall-breaking effect of lysozyme resistant bacteria, the success rate and DNA yield through physical bead grinding.

Advantages

• Fast – several samples can be extracted in 40 minutes (after digestion)
• High purity – purified DNA can be directly used in various downstream applications
• Good repeatability – silica technology can obtain ideal results every time
• High recovery – DNA can be recovered at the level of PG
• Broad spectrum – it can deal with a variety of bacteria, including Gram-positive bacteria that are difficult to disrupt
• Sufficient components – this kit has carried lysozyme, protease K, RNase A and glass beads

Kit Contents

Contents	D314602	D314603
Purification Times	50 Preps	250 Preps
Hipure DNA Mini Columns I	50	2 x 125
2ml Collection Tubes	50	2 x 125
Glass Beads (0.1~0.2mm)	20 g	100 g
Buffer P1	20 ml	100 ml
Buffer DL	15 ml	80 ml
Buffer GW1	22 ml	88 ml
Buffer GW2	12 ml	50 ml
Lysozyme	60 mg	300 mg
Proteinase K	24 mg	120 mg
Protease Dissolve Buffer	5 ml	15 ml
Buffer AE	15 ml	60 ml

Storage and Stability

Lysozyme and Proteinase K should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions.

This product provides fast and easy methods for purification of total DNA for reliable PCR and southern blotting. Total DNA can be purified from bacterial culture, body fluids, food and fermentation.

The Opentrons® 3-piece Aluminum Block Set is used with our Temperature Deck. The set comes with a 24-well, 96-well and flat bottom plate format block. These are used to keep your 2mL tubes, 1.5mL tubes, 96-well PCR plates, PCR strips and flat bottom plates at a constant temperature. These blocks holds temperatures between 4°C and 95°C when used with the Opentrons Temperature Module (GEN2).

The Opentrons® 3-piece Aluminum Block Set is used with our Temperature Deck. The set comes with a 24-well, 96-well and flat bottom plate format block. These are used to keep your 2mL tubes, 1.5mL tubes, 96-well PCR plates, PCR strips and flat bottom plates at a constant temperature. These blocks holds temperatures between 4°C and 95°C when used with the Opentrons Temperature Module (GEN2).

Description 

The PM5200 3-color Pre-Stained Protein Ladder Broad Range is a ready-to-use three-color protein standard with 15 pre-stained proteins covering a wide range of molecular weights from 5 to 245 kDa in Tris-Glycine Buffer (3.5 to 235 kDa in Bis-Tris (MOPS) buffer and Bis-Tris (MES) buffer). Proteins are covalently coupled with different chromophores for easy identification of bands, with three reference proteins carrying enhanced intensity corresponding to a blue band at 20 kDa, green at 40 kDa, and red at 75 kDa, respectively, as separated on SDS-PAGE (Tris-Glycine buffer). The PM5200 3-color Pre-Stained Protein Ladder Broad Range 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.
• Three reference bands — 75 kDa (red), 40 kDa (green), and 20 kDa (blue)

Contents 

Approximately 0.1~0.4 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, PM5200 ExcelBand™ 3-color Pre-Stained Protein Ladder Broad Range resolves 15 major bands in SDS-PAGE (Tris-Glycine, MOPS, and MES buffer) and after Western blotting to nitrocellulose membrane. 

Storage

4°C for 3 months
-20°C for long term storage 

The PM5200 3-color Pre-Stained Protein Ladder Broad Range is a ready-to-use three-color protein standard with 15 pre-stained proteins covering a wide range of molecular weights from 5 to 245 kDa in Tris-Glycine Buffer (3.5 to 235 kDa in Bis-Tris (MOPS) buffer and Bis-Tris (MES) buffer). Proteins are covalently coupled with different chromophores for easy identification of bands, with three reference proteins carrying enhanced intensity corresponding to a blue band at 20 kDa, green at 40 kDa, and red at 75 kDa, respectively, as separated on SDS-PAGE (Tris-Glycine buffer). The PM5200 3-color Pre-Stained Protein Ladder Broad Range 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.