The NGS FFPE DNA Library Prep Kit (illumina and MGI Platforms) was developed for the construction of high quality FFPE DNA libraries using 10 ng to 400 ng of input DNA isolated from formalin-fixed, paraffin-embedded (FFPE) samples. The DNA damage caused by fixation makes it difficult to construct high quality libraries. Our kit has been optimized to repair damaged DNA in the reactions. Multiplexing of the NGS FFPE DNA Library is possible.
NGS FFPE DNA Library Prep Kit Workflow
FFPE samples are a great resource for biomedical research. However, the methods for fixation and condition of storage significantly damage the DNA in the samples. Thus, the extraction of high quality DNA from FFPE samples is often a challenge. Low yield and low quality of FFPE DNA usually are common because of the limited tissue material and the DNA degradation.
As a result, it is usually difficult to construct high quality NGS libraries from low amount and low quality of FFPE DNA. In order to address this issue, we have developed the NGS FFPE DNA Library Prep Kit to make high quality libraries from the low input of FFPE DNA samples.
Three index types are available for the NGS FFPE DNA Library Prep Kit of the illumina platform:
Non-index (Cat.# 30035): Libraries do not have index.
Index (Cat.# 30037): Each of our index primers contains a unique barcode DNA (6 bases long) that can be used to identify individual library. Multiplexing of libraries is up to 48 samples. Index information can be downloaded here.
Unique dual index (Cat.# 30039): FFPE DNA library multiplexing is possible with 96 samples based on the unique dual indexing system. Our unique Four–Base Difference Index System allows us to make indexes that have at least 4 bases different from each other in the 8-base index sequence. Our unique dual indexing primers can effectively remove NGS errors including index hopping, de-multiplexing errors, index cross-contamination, mis-assignments etc. The unique dual index primer set includes 96 pre-mixed unique pairs of i5 and i7 index primers. Index information can be downloaded here.
Indexes are available for the MGI platform kits (Cat.# 34037).
Kit advantages:
Comparison of library conversion efficiency under the same condition. Input DNA amount is 25 ng.
Comparison of library yield under the same condition. Input DNA amount is 25 ng.
The NGS FFPE DNA Library Prep Kit (illumina and MGI Platforms) was developed for the construction of high quality FFPE DNA libraries using 10 ng to 400 ng of input DNA isolated from formalin-fixed, paraffin-embedded (FFPE) samples. The DNA damage caused by fixation makes it difficult to construct high quality libraries. Our kit has been optimized to repair damaged DNA in the reactions. Multiplexing of the NGS FFPE DNA Library is possible.
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
The FastRunner DNA Ladder is prepared to ensure quality and batch-to-batch consistency. This Ladder contains eight discrete fragments ranging from 50 bp to 2000 bp with a higher intensity reference band at 500 bp. This Ladder is ideal for quick sizing of PCR products and restriction digests.
Contents
1mL of premixed DNA ladder (0.5µg/10µL) in loading buffer (10mM EDTA, 10% glycerol, 0.015% bromophenol blue, and 0.17% SDS).
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FastRunner DNA Ladder (Cat# 12800) – 100 loads
Ladder Properties:
• Eight discrete bands, ranging from 50 bp to 2000 bp
• Higher intensity band at 500 bp for easy reference
| Fragment | Size (bp) | Mass (ng) |
| 1 | 2000 | 104 |
| 2 | 1500 | 88 |
| 3 | 1000 | 68 |
| 4 | 750 | 59 |
| 5 | 500 | 93 |
| 6 | 300 | 28 |
| 7 | 150 | 35 |
| 8 | 50 | 25 |
Recommended Use:
Mix thoroughly. For best results, load 10µL of DNA ladder per well. For precise mass determination with a densitometer, stain gel after electrophoresis using 0.5µg/mL ethidium bromide for 30-40 minutes. The table above shows the size and mass for each band based on 10µL ladder per well.
Storage:
Stable at room temperature. For longer term storage, -20°C is recommended.
This ladder was standardized using 10µL of DNA per lane on a 0.8 cm thick, 13 x 15 cm, 1.0% agarose gel run in TAE buffer.