N-(t-butyl ester-PEG4)-N-bis(PEG4-Propargyl)

Overview

• Detection kits for the BKV/JCV
• Available in TaqMan format for analysis

The BK virus is a member of the polyomavirus family. It has been suggested that this virus may be transmitted through respiratory fluids or urine, since infected individuals periodically excrete virus in the urine. BK viral infections are typically asymptomatic in healthy individuals, however very mild symptoms may appear including mild respiratory infections and fever. Infections with BK virus in immunocompromised or immunosupressed patients are much more severe and may involve renal dysfunction. In fact, in kidney transplant patients the immunosupressive drugs required for the transplant may allow the virus to replicate within the graft, resulting in a disease called BK virus nephropathy (BKVN). The JC virus is a type of human polyomavirus and is very common in the general population, infecting 70 to 90% of humans. Most people acquire JCV in childhood or adolescence. Typically the infection is subclinical and no of consequence in individuals with healthy immune systems. The initial site of infection may be the tonsils or the gastrointestinal tract, and the virus then remains latent in the gastrointestinal tract. JCV can also infect the tubular epithelial cells in the kidneys, where it continues to reproduce, shedding virus particles in the urine. Also, JCV can cross the blood-brain barrier into the central nervous system. JCV is known to cause the usually fatal progressive multifocal leukoencephalopathy (PML) by destroying oligodendrocytes in the brain in immunodeficient or immunosuppressed individuals. The JC and BK viruses are very similar, with their genomes sharing 75% homology. It is however important to differentiate between the viruses due to the differences in pathology and especially the invariably fatal outcome of PML which is only caused by the JC virus.

Details

Supporting Data

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Storage Conditions and Product Stability
All kit components can be stored for 2 years after the date of production without showing any reduction in performance.

All kit components should be stored at -20°C upon arrival.

Component	Cat. TM39350 (100 preps)	Cat. TM39310 (100 preps)
MDx TaqMan 2X PCR Master Mix	2 x 700 μL	–
BKV/JCV Primer & Probe Mix	280 μL	280 μL
BKV/JCV Positive Control	150 μL	150 μL
Nuclease-Free Water (Negative Control)	1.25 mL	1.25 mL
Product Insert	1	1

DBCO-PEG12-acid is an analog of DBCO-Acid with hydrophilic PEG linker and a DBCO group. The DBCO groups is commonly used for Click Chemistry reactions. The hydrophilic PEG chain allows for increased water solubility of compounds in aqueous media. The terminal carboxylic acid can react with primary amine groups in the presence of activators (e.g. EDC, or HATU) to form a stable amide bond. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

DBCO-PEG12-acid is an analog of DBCO-Acid with hydrophilic PEG linker and a DBCO group. The DBCO groups is commonly used for Click Chemistry reactions. The hydrophilic PEG chain allows for increased water solubility of compounds in aqueous media. The terminal carboxylic acid can react with primary amine groups in the presence of activators (e.g. EDC, or HATU) to form a stable amide bond. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

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 V3-V4 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V3-V4 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 V3-V4 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 V3-V4 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

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 V3-V4 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. 70400 (24 preps)	Cat. 70410, 70420, 70430, 70440 (96 preps)
Amplicon PCR (PCR 1)	MGX Master Mix	330 µL	1,320 µL
	16S V3-V4 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