GeneAb™ PD-L1

The NGS Low Input DNA Library Prep Kit (illumina and MGI Platforms) was developed for construction of high quality NGS libraries with low input DNA amount from 1 ng to 400 ng. The kit allows scientist to study samples with limited DNA such as tumor samples, patient samples, and other specially collected samples (FACS sorting etc.). The kit has high library conversion efficiency with as little as 1 ng DNA input. The fast and simple 1.5-hour protocol makes libraries with even coverage and low GC-bias based our unique chemistry for DNA end-polishing and ligation.

The kit needs double strand DNA fragments (blunt and/or sticky) as input DNA for NGS library preparation, and is compatible with DNA fragments generated from both enzymatic approach (for example, BioDynami DNA fragmentation enzymes, Cat.# 40061 and Cat.# 40062) and mechanical approaches such as sonication and nebulization etc.

NGS Low Input DNA Library Prep Kit Workflow

Three index types are available for the NGS Low Input DNA Library Prep Kit of illumina platform:

Non-index (Cat.# 30022): Libraries do not have index.

Index (Cat.# 30024): Each of our index primers contains a unique barcode sequence with 6 bases that can be used to identify the low input DNA libraries. Library multiplexing up to 48 samples is possible. Index information can be downloaded here.

Unique dual index (Cat.# 30025): Library multiplexing up to 96 low input DNA libraries is possible with unique dual indexes with our Four-Base Difference Index System. The system allows us to make indexes for the libraries that have at least 4 bases different from each other in the 8-base barcode length. Our unique dual index primers can reduce sequencing errors such as de-multiplexing errors, amplification errors, mis-assignment of reads, index cross-contamination, and also index hopping. The kit  includes 96 pre-mixed unique pairs of index primers. Index information can be downloaded here.

Indexes are available for the MGI platform kits (Cat.# 34024).

Kit advantages:

• • • Fast protocol
      • The hands-on time is only 10 minutes
      • The total protocol time is around 1.5 hours
    • Simple procedure
      • Ready-to-use master mix makes it simple for reaction setup
      • Less reaction components
    • Less magnetic beads required for cleanup steps: Save the cost more than 50%
    • Low input DNA amount: Starts from 1 ng of DNA

Comparison of library conversion efficiency under the same NGS library preparation condition. Input DNA amounts are 1 ng and 10 ng, respectively. DNA was mechanical sheared with Covaris before library prep. BioDynami kit: NGS Low Input DNA Library Prep Kit (Cat. #30022).

Comparison of library yield under the same NGS library prep condition. Input DNA amounts are 1 ng, 10 ng, and 100 ng. DNA was mechanical sheared with Covaris before library prep. BioDynami kit (Cat. #30022). PCR cycle numbers were indicated.

Overview

• Detection kits for Chlamydia/Neisseria gonorrhoeae
• Available in TaqMan format for analysis

Chlamydia trachomatis, the causative agent of Chlamydia, is a Gram negative bacteria. Transmission of the bacteria occurs via contact with infected bodily fluids which then infect mucosal membranes. It can be transmitted from mother to child during pregnancy and infect the eyes causing conjunctivitis. The genital infection causes urethritis, epididymitis and prostatitis in males and Pelvic Inflammatory Disease (PID) in females with an increased risk of contracting HIV. The infection can be treated with a course of antibiotics. Sexually transmitted infections in females are most often asymptomatic, but can be noticeable in chronic pain of the pelvic region, vaginal bleeding and painful urination. Infection of the ovaries, fallopian tubes or uterus causes Pelvic Inflammatory Disease (PID) which can lead to difficulties in conceiving, increased risk of ectopic pregnancy or infertility. Infections in males are more likely to be symptomatic, causing painful urination, discharge from the penis and swollen testicles and may eventually cause infertility if left untreated.

Neisseria gonorrhoeae is a Gram-negative coccus of the Neisseria genus. N. gonorrhoeae is usually seen in pairs infecting human cells. It has a circular DNA genome of approximately 1Mbp encoding over 2000 genes. N. gonorrhoeae is transmitted by sexual contact and usually causes infection in cells of the mucous membrane of the male urethra or the endocervix and urethra in females. During infection, polysaccharides are released from the bacteria that stimulate host cell production of tumour necrosis factors that cause an inflammatory response. There is no vaccine against N. gonorrhoeae infection and antibiotic resistance is beginning to increase, therefore treatment includes a course of antibiotics that will be effective against resistant strains. Complications in males caused by the infection can result in prostatitis or orchitis if the bacteria spread. In females, invasion of the fallopian tubes or ovaries can result in salpingitis or ovaritis respectively, with any of these infections possibly resulting in sterility.

Chlamydia/Neisseria TaqMan PCR Kit,
100 reactions

• Ready to use format, including Master Mix for the target and PCR control to monitor for PCR inhibition and validate the quality
• Specific Primer and Probe mix for the pathogen/virus/viroid of interest
• Primer and Probe mix
• Positive and negative control to confirm the integrity of the kit reagents

Chlamydia/Neisseria TaqMan PCR Probe/Primer Set and Controls,
100 reactions

• Specific Primer/Probe mix and Positive Control for the pathogen/virus/viroid of interest
• Nuclease-free water
• Can be used together with Norgen’s PCR Master Mix (#28007) or customer supplied master mix

For research use only and NOT intended for in vitro diagnostics.

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. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.

Component	Cat. TM42550 (100 preps)	Cat. TM42510 (100 preps)
MDx TaqMan 2X PCR Master Mix	2 x 700 μL	–
Chlamydia/Neisseria Primer & Probe Mix	280 μL	280 μL
Chlamydia/Neisseria Positive Control	150 μL	150 μL
Nuclease-Free Water (Negative Control)	1.25 mL	1.25 mL
Product Insert	1	1

Description 

The ExcelDye™ 6× DNA Loading Dye (Tri-Color) is pre-mixed buffer for tracking the DNA sample during the electrophoresis on agarose or polyacrylamide gels. It contains three dyes (Xylene cyanol FF, Bromophenol blue, Orange G) for tracking the DNA migration. The Xylene cyanol FF, Bromophenol blue and Orange G migrate at approximately 800 bp, 150 bp and 30 bp on a standard 2% TAE agarose gel respectively (4,000 bp, 500 bp and 50 bp on 1% TAE agarose gel respectively). The included glycerol keeps the DNA at the bottom of the well and the presence of EDTA chelates divalent metal ions to prevent the process of metal-dependent nuclease. 

Composition 

• 0.03% Xylene cyanol FF 
• 0.03% Bromophenol blue
• 0.15% Orange G 
• 10 mM Tris-HCl (pH 8.0) 
• 60% glycerol 
• 60 mM EDTA

Storage 

4°C for 12 months
-20°C for 36 months

The ExcelDye™ 6× DNA Loading Dye (Tri-Color) is pre-mixed buffer for tracking the DNA sample during the electrophoresis on agarose or polyacrylamide gels. It contains three dyes (Xylene cyanol FF, Bromophenol blue, Orange G) for tracking the DNA migration. The Xylene cyanol FF, Bromophenol blue and Orange G migrate at approximately 800 bp, 150 bp and 30 bp on a standard 2% TAE agarose gel respectively (4,000 bp, 500 bp and 50 bp on 1% TAE agarose gel respectively). The included glycerol keeps the DNA at the bottom of the well and the presence of EDTA chelates divalent metal ions to prevent the process of metal-dependent nuclease.