Human Immunodeficiency Virus (HIV) is a retrovirus which destroys essential cells in the human immune system, such as CD4+ lymphocytes (a type of T cell), macrophages and dendritic cells. Infection with HIV therefore leads to an impaired immune system and increased susceptibility to opportunistic cancers and infections. Due to the mutating nature of HIV and damage to the T cells, the immune system is unable to fight off infections and disease. Without treatment, HIV can progress to Acquired Immunodeficiency Syndrome (AIDS). The rate of HIV progression to AIDS depends on a number of different factors including viral, host and environmental factors. However, with adherence to antiretroviral therapy (ART), many individuals with HIV will never progress to AIDS, having near normal life expectancy. Transmission of HIV is through direct contact with infected bodily fluids, either sexually; by the sharing of blood-contaminated needles or instruments; maternally through childbirth or breastfeeding; or via blood transfusion/organ transplant.
HIV TaqMan RT-PCR Kit, 100 reactions
HIV TaqMan RT-PCR Probe/Primer Set and Controls, 100 reactions
For research use only and NOT intended for in vitro diagnostics.
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Storage Conditions and Product Stability
All kit components can be stored for 1 year 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. TM33750 (100 preps) | Cat. TM33710 (100 preps) |
|---|---|---|
| MDx TaqMan 2X PCR Master Mix | 2 x 700 μL | – |
| HIV Primer & Probe Mix | 280 μL | 280 μL |
| HIV Positive Control | 150 μL | 150 μL |
| Nuclease-Free Water (Negative Control) | 1.25 mL | 1.25 mL |
| Product Insert | 1 | 1 |
The PCR-free NGS DNA Library Prep Kit was developed for construction of DNA libraries for next generation sequencing (illumina platform). The kit adds 3′-dT-tailed library adapters to both ends of DNA fragments efficiently. The kit uses double strand DNA fragments (blunt and/or sticky) as input DNA for NGS library construction, and is compatible with DNA fragments generated from both enzymatic methods (BioDynami DNA fragmentation enzymes etc.) and physical methods (sonication, nebulization etc.).
PCR-Free NGS DNA Library Prep Kit Workflow
PCR amplification is a standard step for library preparation of Next-Generation Sequencing. The PCR is used to amplify fully ligated DNA fragments and to add index information to the libraries. The indexing is for the pooling of the library samples in an effort to reduce the sequencing cost.
However, PCR introduces uneven amplification of DNA in some DNA regions with extreme GC-contents and secondary structures. This bias can cause very low sequencing coverage in such regions. DNA sequencing in these regions is still a huge challenge.
PCR-free library prep can reduce library bias and minimize sequencing gaps. The sequencing data from PCR-free library samples have even genomic coverage with few gaps and better depth in GC-rich regions. Our PCR-free NGS kit offers optimal coverage in the regions that are traditionally difficult, such as high-GC content regions, low-GC content regions, and repetitive sequence regions.
Two index types are available for the kit:
Non-index: Libraries do not have index.
Index: Each library contains one i5 index and one i7 index. Library multiplexing up to 96 samples is possible. List of indexes can be downloaded Here.
Kit advantages:
The PCR-free NGS DNA Library Prep Kit was developed for construction of DNA libraries for next generation sequencing (illumina platform). The kit adds 3′-dT-tailed library adapters to both ends of DNA fragments efficiently. The kit uses double strand DNA fragments (blunt and/or sticky) as input DNA for NGS library construction, and is compatible with DNA fragments generated from both enzymatic methods (BioDynami DNA fragmentation enzymes etc.) and physical methods (sonication, nebulization etc.).
These kits provide everything required to isolate and purify inclusion body proteins from induced bacterial cultures. First a proprietary Cell Lysis Reagent is used to selectively lyse the cells and release inclusion bodies in their solid form. Next, inclusion bodies are dissolved and their contents released using the provided IB Solubilization Reagent. Inclusion body proteins are then further purified using spin columns for rapid and convenient buffer exchange and desalting. This kit provides a convenient way to screen recombinants prior to scaling up.
ProteoSpin™ Inclusion Body Protein Isolation Micro Kit
The process is efficient and streamlined and can process up to 12 samples in only 60 minutes. Each spin column is able to recover up to 50 µg of acidic or basic proteins. Purified recombinant proteins are then ready for SDS-PAGE, 2D gels, Western blots, Mass Spectrometry analysis, and other applications.
ProteoSpin™ Inclusion Body Protein Isolation Maxi Kit
The procedure is efficient and streamlined and can process up to 4 samples in approximately 2 hours. Each spin column is able to recover up to 12 mg of acidic or basic proteins from 100 mL of induced bacterial culture. Purified recombinant proteins are then ready for SDS-PAGE, 2D gels, Western blots, Mass Spectrometry analysis, and other applications.
About Inclusion Bodies
Bacteria are widely used for the expression of different proteins. However, 70-80% of the proteins expressed in bacteria by recombinant techniques are typically contained in insoluble inclusion bodies (i.e., protein aggregates). The protein of interest found in these subcellular structures is often inactive, due to incorrect folding. The production rate of recombinant proteins stored in inclusion bodies is invariably higher than those synthesized as soluble proteins. The reason behind this is thought to be the resistance of insoluble proteins to proteolysis by cellular enzymes. In addition, separation of insoluble recombinant proteins in inclusion bodies is considerably easier than that of soluble proteins. These factors have been the major influences favoring scale-up of high-value proteins using bacterial fermentation for example. Procedures for the purification of the expressed proteins from inclusion bodies are often labour-intensive, time-consuming and not cost-effective. This kit provides the essential reagents for cell disruption, inclusion body solubilization and purification using spin column chromatography – all optimized to work together thereby simplifying the process and saving a tremendous amount of time and cost.
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| Kit Specifications | |
| Maximum Culture Volume | 100 mL |
| Yield from 100 mL of Culture | Up to 12 mg |
| Minimum Elution Volume | 4 mL |
| Time to Process 1 Sample | 1 hour |
Storage Conditions
The Cell Lysis Reagent and IB Solubilization Reagent should be stored at 4°C upon receipt of this kit. This kit is stable for 2 years after the date of shipment. Once opened, the solutions should be stored at 4°C when not in use except for Binding Buffer C and Binding Buffer N. Some precipitation will occur with 4°C storage. This precipitation should be dissolved with slight heating to room temperature before using.
| Component | Cat. 10300 (Micro – 25 preps) | Cat. 17700 (Maxi – 4 preps) |
|---|---|---|
| Wash Solution C | 30 mL | 130 mL |
| Wash Solution N | 30 mL | 130 mL |
| Binding BUffer A | 4 mL | 20 mL |
| Binding Buffer N | 4 mL | 20 mL |
| Elution Buffer C | 8 mL | 2 x 30 mL |
| Protein Neutralizer | 4 mL | 4 mL |
| Cell Lysis Reagent | 15 mL | 110 mL |
| IB Solubilization Reagent | 2 mL | 50 mL |
| Syringes, 1cc, slip tip | 25 | – |
| Needles (Bev, 20G x 1 inch) | 25 | – |
| Syringes, 10 mL, Luer-Lok™ Tip | – | 4 |
| Needles (18G x 1.5 inch) | – | 4 |
| Micro Spin Columns | 25 | – |
| Maxi Spin Columns (filled with SiC) inserted into 50 mL collection tubes | – | 4 |
| Collection Tubes | 25 | – |
| Elution Tubes (1.7 mL) | 25 | – |
| Elution Tubes (50 mL) | – | 4 |
| Product Insert | 1 | 1 |