ExcelRT™ Reverse Transcription Kit is a complete, efficient and convenient kit to synthesize high quality first strand cDNA. This kit contains ExcelRT™ Reverse Transcriptase, which is able to synthesize the first strand cDNA at 37~50°C. The ExcelRT™ Reverse Transcriptase is a recombinant Moloney Murine Leukemia Virus (M-MLV) reverse transcriptase, which is designed to reduce RNase H activity and create better thermal stability. This kit also contains RNAok™ RNase Inhibitor, which is active against RNase A, RNase B, and RNase C. This product is supplied with oligo (dT)20 and random hexamers, which are used to synthesize cDNA from poly(A) tailed mRNA and total RNA, respectively.
Detail
Description
ExcelRT™ Reverse Transcription Kit is a complete, efficient and convenient kit to synthesize high quality first strand cDNA. This kit contains ExcelRT™ Reverse Transcriptase, which is able to synthesize the first strand cDNA at 37~50°C. The ExcelRT™ Reverse Transcriptase is a recombinant Moloney Murine Leukemia Virus (M-MLV) reverse transcriptase, which is designed to reduce RNase H activity and create better thermal stability. This kit also contains RNAok™ RNase Inhibitor, which is active against RNase A, RNase B, and RNase C. This product is supplied with oligo (dT)20 and random hexamers, which are used to synthesize cDNA from poly(A) tailed mRNA and total RNA, respectively.
Features
Contains all components for reverse transcription
High yield
Thermostable, up to 50°C, during first strand synthesis
High processivity, generating cDNA up to 8 kb
Reduced RNase H ribonuclease activity
Application
Generation of first strand cDNA from total RNA or mRNA.
Suitable for generating cDNA from RNA with strong secondary structure which can be reduced at temperature up to 50°C.
Storage
-20°C for 24 months
High yield
High sensitivity
Thermostable, up to 50°C, during first strand synthesis
Contents
ComponentVolume ExcelRT™ Reverse Transcriptase (200 U/μl) 100 μl RNase Inhibitor (20 U/μl)100 μl 5X RT Buffer (DTT)500 μl dNTPs Mix (10 mM each)200 μl Oligo (dT)20 (50 μM)100 μl Random Hexamers (100 μM)100 μl DEPC-Treated H2O 1 ml x 2
Storage Buffer
Reverse Transcriptase: 20 mM Tris-HCl (pH 7.5), 200 mM NaCl, 0.1 mM EDTA, 1 mM DTT, stabilizer and 50% (v/v) glycerol
RNase Inhibitor: 40 mM HEPES-KOH (pH 7.5), 100 mM KCl, 8 mM DTT, 0.1 mM EDTA, stabilizer and 50% (v/v) glycerol
5X RT buffer (DTT)
250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM MgCl2 and 50 mM DTT
CE-IVD marked version available for in vitro diagnostic use
Available in TaqMan format for analysis
Herpes Simplex Virus 2 (HSV-2) is a member of the herpes virus family, Herpesviridae. HSV-2 has a relatively large double-stranded DNA genome. HSVs are primarily transmitted by sexual intercourse, direct contact with lesions or perinatally. Most HSV positive cases are characterised by lesions on the skins and mucous membranes of the mouth and genitals. HSV infection can be either primary or a recurrence of a previous infection. More than 90% of the primary HSV infections are asymptomatic. Primary infection with HSV-1 can lead to gingivostomatitis, eczema herpeticum, keratoconjunctivitis and encephalitis. The primary symptoms of a secondary infection are skin lesions in the nose, mouth and genital regions. The infection is contagious, mainly during an epidemic.
HSV-2 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
HSV-2 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
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.
All-in-one solution for inclusion body protein isolation and purification
Fast and convenient spin column protocol
Complete kit with Cell Lysis Reagent, Inclusion Body Solubilization Reagent, buffers and spin columns to purify proteins
Purification is based on spin column chromatography that uses Norgen’s resin separation matrix
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.
The Cell Lysis Reagent and IB Solubilization Reagent should be stored at 4°C upon receipt of this kit. All other solutions should be kept tightly sealed and stored at room temperature. 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 the pH Binding Buffers (Acidic and Basic). 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
