Clostridium difficile is rod-shaped, gram positive bacterium. It is the main causal agent of antibiotic-associated diarrhea and pseudomembranous colitis. The colonization of intestines by C. difficile is usually associated with the elimination of natural intestinal flora as a result of antibiotic application and is frequently reported in health care centers. While C. difficile infection could be severe and life-threatening, particularly among the elderly, many patients are asymptomatic making diagnosis challenging during outbreaks. The tradition method of detecting C. difficile infection is by cytotoxicity test of the toxin produced by the bacterium, but such protocols usually require extensive time before conclusion can be made.
Figure 1 / 3
Click for expanded view
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. TM37150 (100 preps) | Cat. TM37110 (100 preps) |
|---|---|---|
| MDx TaqMan 2X PCR Master Mix | 2 x 700 μL | – |
| Clostridium difficile Primer & Probe Mix | 280 μL | 280 μL |
| Clostridium difficile Positive Control | 150 μL | 150 μL |
| Nuclease-Free Water (Negative Control) | 1.25 mL | 1.25 mL |
| Product Insert | 1 | 1 |
Mal-amido-PEG4-alkyne is a short PEG linker featuring a maleimide and a terminal alkyne. Maleimide is a covalent ligand used for linking thiols such as those in cysteines residues in proteins, while terminal alkynes can be used in copper (I) click chemistry with azides on target molecules.
Mal-amido-PEG4-alkyne is a short PEG linker featuring a maleimide and a terminal alkyne. Maleimide is a covalent ligand used for linking thiols such as those in cysteines residues in proteins, while terminal alkynes can be used in copper (I) click chemistry with azides on target molecules.
Sample type purification kit guide
The 16S V1-V3 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V1-V3 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-V3 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-V3 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.
Figure 1 / 3
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-V3 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. 70200 (24 preps) | Cat. 70210, 70220, 70230, 70240 (96 preps) |
|---|---|---|---|
| Amplicon PCR (PCR 1) | MGX Master Mix | 330 µL | 1,320 µL |
| 16S V1-V3 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 |