Sample type purification kit guide
The ITS2 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the fungal target ITS2 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 XPMagneticBeads(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 ITS2 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 ITS2 region of the fungal DNA. The post-PCR reaction is then cleaned up using AMPure XPbeads. 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 XPbeads. The libraries are then ready for quantification, pooling and sequencing.
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| 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 ITS2 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 will remain stable for at least 1 year when stored at the specified storage conditions.
K-PullG6
SKU: 700004329
100/200 assays per kit
| Content: | 100 / 200 assays per kit |
| Shipping Temperature: | Ambient |
| Storage Temperature: | Short term stability: 2-8oC, Long term stability: See individual component labels |
| Stability: | > 2 years under recommended storage conditions |
| Analyte: | Pullulanase/Limit-Dextrinase |
| Assay Format: | Spectrophotometer |
| Detection Method: | Absorbance |
| Wavelength (nm): | 400 |
| Signal Response: | Increase |
| Limit of Detection: | 0.18 U/mL for pullulanase preparations (50-fold dilution) 0.01 U/g for limit dextrinase in milled malt |
| Reproducibility (%): | ~ 3% |
| Total Assay Time: | ~ 10 min (Pullanase), ~ 30 min (Limit-Dextrinase) |
| Application examples: | Assay of microbial pullulanase preparations. Measurement of limit-dextrinase in malt extracts. |
| Method recognition: | Novel method |
PullG6 assay for the measurement of pullulanase employs a water soluble defined substrate, namely 4,6-O-benzylidene-4-nitrophenyl-63-α-D-maltotriosyl-maltotriose (BPNPG3G3), coupled with the ancillary enzymes α-glucosidase and β-glucosidase. Upon hydrolysis of the substrate at the 1,6-α-linkage by pullulanase or limit-dextrinase, the released 4-nitrophenyl-β-maltotrioside is immediately hydrolysed to glucose and 4-nitrophenol by the concerted action of the α-glucosidase and β-glucosidase enzymes in the reagent mixture. The reaction is terminated and phenolate ions are developed by addition of dilute alkali. The absorbance is read at 400 nm and the value obtained correlates directly with pullulanase activity.
Explore more of our assay kit products for enzyme activity measurement.
Advantages
DBCO-PEG12-amine is a PEG linker which contains DBCO and amine moieties. The DBCO group is commonly used in copper-free Click Chemistry reactions. The amine group is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde), etc. The hydrophilic PEG spacer increases the water solubility of the compound. Reagent grade, for research purpose.
DBCO-PEG12-amine is a PEG linker which contains DBCO and amine moieties. The DBCO group is commonly used in copper-free Click Chemistry reactions. The amine group is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde), etc. The hydrophilic PEG spacer increases the water solubility of the compound. Reagent grade, for research purpose.