12x8ml PCR Rotor 

Description 

The ExcelDye™ 6× DNA Loading Dye (Orange) is pre-mixed buffer for tracking the DNA sample during the electrophoresis on agarose or polyacrylamide gels. It contains one dye (Orange G) for tracking the DNA migration. The Orange G migrates at approximately 30 bp on a standard 2% TAE agarose gel (50 bp on 1% TAE agarose gel). The included glycerol keeps the DNA at the bottom of the well and the presence of EDTA chelates divalent metal ions to prevent the process of metal-dependent nuclease.

Composition 

• 0.15% Orange G 
• 10 mM Tris-HCl (pH 8.0) 
• 60% glycerol 
• 60 mM EDTA

Storage 

4°C for 12 months
-20°C for 36 months

The ExcelDye™ 6× DNA Loading Dye (Orange) is pre-mixed buffer for tracking the DNA sample during the electrophoresis on agarose or polyacrylamide gels. It contains one dye (Orange G) for tracking the DNA migration. The Orange G migrates at approximately 30 bp on a standard 2% TAE agarose gel (50 bp on 1% TAE agarose gel). The included glycerol keeps the DNA at the bottom of the well and the presence of EDTA chelates divalent metal ions to prevent the process of metal-dependent nuclease.

Overview

• Efficient depletion of bovine exosomes from Fetal Bovine Serum
• Deplete exosome-sized vesicles from versatile FBS volumes
• No protease treatment required
• No time-consuming ultracentrifugation
• No precipitation reagents required
• No overnight incubation required
• Exosome depletion confirmed by reduction of bovine miRNAs below detectable levels
• The depleted FBS provides the same cellular growth rates as the standard FBS
• Purification is based on Norgen’s proprietary Silicon Carbide resin matrix

Norgen’s FBS Exosome Depletion Kits provides a quick and easy protocol for the depletion of bovine exosomes from FBS prior to using it as a growth supplement in your culture medium. The FBS recovered from the depletion process is exosome-depleted and does not contain any quantifiable bovine miRNAs. Moreover, the exosome-depleted FBS will support the growth of your cells of interest similar to the non-depleted FBS. Norgen’s kits allow for the processing of different FBS volumes. The depletion is based on Norgen’s proprietary resin. These kits provide a clear advantage over other available kits in that they do not require ultracentrifugation, any special instrumentation, precipitation reagents or any protease treatments. More importantly, the depletion process is an inexpensive method for exosome depletion from FBS, as compared to the expensive current ready-to-use exosome-depleted media available on the market.

Background

Most culture medium used for the growth and propagation of cells in culture require the addition of fetal bovine serum (FBS) as a growth supplement to media. FBS is obtained from bovine (cow) serum, and therefore contains large quantities of bovine exosome vesicles. These exosomes may interfere with some types of studies, or may lead to unreliable results when studying the exosomes shed from your cells of interest in normal culture conditions. Therefore, the use of exosome-depleted FBS is highly recommended for many types of studies.

FBS Exosome Depletion Kit (Slurry Format)

For FBS volumes ranging from 140 mL to 280 mL.

FBS Exosome Depletion Kit (Column Format)

For FBS volumes ranging from 120 mL to 240 mL.

Details

Supporting Data

Figure 1 / 3

Previous

Next

Click for expanded view

Kit Specifications
Sample Type	Fetal Bovine Serum
Sample Volume Range	Up to 140 mL (FBS Exosome Depletion Kit I (Slurry Format) Up to 280 mL (FBS Exosome Depletion Kit II (Slurry Format)
Depletion	Deplete exosome-sized vesicle
Bovine miRNA	No detectable bovine miRNA
Time to Complete 6 Purifications	40 minutes

Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.

Component	Cat. 61100 (6 preps)	Cat. 61400 (12 preps)	Cat. 61200 (6 preps)	Cat. 61300 (12 preps)
ExoC Buffer	2 x 1.5 mL	8 mL	2 x 1.5 mL	8 mL
Slurry E	12.5 mL	2 x 12.5 mL	–	–
Maxi Spin Column	–	–	6	12
Product Insert	1	1	1	1

Overview

• Protocol optimized for DNA isolated from a diversity of samples including stool, soil, water, saliva, plant, urine, skin, and more
• Simple and quick workflow: library could be prepared in less than 5 hours
• Component of Norgen’s metagenomics workflow
• A single NGS run can be prepared with up to 384 unique dual-index libraries

Sample type purification kit guide 

The 16S V1-V2 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V1-V2 amplicon libraries to be used for next-generation sequencing on Illumina platforms. All molecular reagents including primers, enzyme mixes, indexes, and buffers are provided. Instructions for PCR clean up with the AMPure XP Magnetic Beads (supplied by customer) are also included for rapid purification of nucleic acid products generated at two steps of the workflow. The library prep workflow could be used for purified DNA inputs from different sources including stool, soil, water, saliva, plant, urine, skin swab, vaginal swab, cheek swab, nasal swab, plasma/serum, tongue swab, gum swab, and others.

The 16S V1-V2 Library Preparation Kit for Illumina has a streamlined procedure that reduces the handling time such that the library prep procedure can be completed in approximately 4 hours (see diagram below). Input DNA is first subjected to targeted PCR to amplify the V1-V2 region of the DNA encoding 16S rRNA. The post-PCR reaction is then cleaned up using AMPure XP beads. Dual index primers are then added using a limited-cycle PCR. The indexed amplicons flanked by 5′ and 3′ barcoded adaptors are then cleaned using AMPure XP beads. The libraries are then ready for quantification, pooling and sequencing.

Details

Supporting Data

Figure 1 / 3

Previous

Next

Click for expanded view

Minimum amount of starting material:	2.5 µL of DNA (5 ng/µL)
Time to complete library preparation:	4 hours

Storage Conditions and Product Stability
Norgen’s 16S V1-V2 Library Prep Kit for Illumina is shipped as one kit box (for the 24 prep kit) or two sub-component kits (for the 96 prep kit). All kits should be stored at -20°C upon arrival.

All kit components should remain stable for at least 1 year when stored at the specified storage conditions.

Step	Component	Cat. 70100 (24 preps)	Cat. 70110, 70120, 70130, 70140 (96 preps)
Amplicon PCR (PCR 1)	MGX Master Mix	330 µL	1,320 µL
	16S V1-V2 Primer Mix	70 µL	280 µL
Index PCR (PCR 2)	Indexing Master Mix	660 µL	2 x 1,320 µL
	N7xx Index Primer	50 µL	50 µL
	S5xx Index Primer	70 µL	70 µL
PCR Clean-Up	Resuspension Buffer	2 x 1,250 µL	2 x 5,000 µL
	Nuclease-free water	1,250 µL	1 x 6,000 µL