Grey, standard profile semi-skirted PCR plate with a rigid two-component hardshell design which eliminates warping and distortion during PCR testing.
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.
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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 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 |
Usages：
Determination of coliform and fecal coliform for multiple tube fermentation.
Principle:
Tryptone provide carbon and nitrogen sources to meet the needs of bacterial growth; sodium chloride osmotic pressure balance can be maintained; Lactose is a coliform fermentable sugars; potassium dihydrogen phosphate and dipotassium phosphate is a buffer; lauryl sodium can inhibit the growth of non-coliform bacteria.
Formulation (per liter):
Trypticase:20g
Sodium chloride: 5g
Lactose:5g
Potassium dihydrogen phosphate: 2.75g
Dipotassium hydrogen phosphate :2.75g
Sodium lauryl sulfate 0.1g
Final pH6.8 ± 0.2
How to use:
1. Suspend 35.6g of the product, adding 1 L of distilled or deionized water, heated to boiling stirring until completely dissolved, packed in a test tube with a small down tube, 121 ℃ autoclave 15min, leave to cool to room temperature, standby .
2.Sample handling and dilution.
3.Selected three consecutive dilution, each dilution was inoculated three LST broth tubes, each tube was inoculated 1mL.
4. Put the tubes in an incubator 36 ± 1 ℃ cultured for 48 ± 2h.
5. Observe the results. if all LST broth don’t pruduse gas, can be reported as negative for E. coli, if gas production then will have to make further confirmed by experiments.
Quality control:
Quality control strains were inoculated and culuture at 36 ± 1 ℃ for 24h ,results are as follows:
Bacterial Name Bacterial No. Growth Status Gassing
Escherichia coli ATCC25922 good +
Salmonella typhimurium CMCC (B) 50115 good —
Staphylococcus aureus ATCC6538 inhibited —
Storage: Store in a dark, cool and dry place, tighten the cap immediately after use. Storage period of three years.
Specifications: 250g/bottle
250g