The GetClone™ PCR Cloning Vector II is a positive selection system for high efficiency cloning of blunt end DNA or amplicons. This cloning vector contains a lethal gene which can be disrupted by ligation of a blunt end DNA insert into the cloning site. Only colonies with inserted vectors are able to propagate, eliminating the additional needs of IPTG and X-Gal for blue/white screening. This cloning vector includes ampicillin and kanamycin resistance genes that can meet the needs of most users.
Detail
Description
The GetClone™ PCR Cloning Vector II is a positive selection system for high efficiency cloning of blunt end DNA or amplicons. This cloning vector contains a lethal gene which can be disrupted by ligation of a blunt end DNA insert into the cloning site. Only colonies with inserted vectors are able to propagate, eliminating the additional needs of IPTG and X-Gal for blue/white screening. This cloning vector includes ampicillin and kanamycin resistance genes that can meet the needs of most users.
Features
Cloning efficiency greater than 90%
IPTG and X-Gal are not required
Accepts a wide range of insert/vector ratios 0.5:1 to 12:1
Accepts insert size from 6 bp to 11 kb
The phosphorylation of PCR fragments is not required
Accepts blunt end amplicon or DNA fragment (not for sticky ends)
The ExcelTaq™ 5X Fluorescent PCR Master Mix is a ready-to-use mixture for amplifying targeted DNA fragments. It is designed to serve as a master mix for virtually all PCR applications. This product is supplied as a 5X concentrated mixture containing all the essential ingredients for PCR with the exception of templates and primers. In addition, the mixture contains electrophoresis tracking dye (Bromophenol blue) and a safer fluorescent DNA staining dye, which enables the user to track the electrophoresis process in real time as well as eliminates the need for the staining process. The resultant PCR reaction mixture is sufficiently dense enough to be loaded directly into a TAE or TBE buffered gel for electrophoresis.
Features
5’→3’ DNA polymerase activity
No detectable 3’→5′ exonuclease (proofreading) activity
Generates PCR products with 3′-dA overhangs
High yield PCR
High reproducibility
Reduced pipetting errors
For direct loading into electrophoresis analysis
DNA bands can be visualized directly under UV or 470 nm blue light illumination
Applications
Routine PCR
Colony PCR
High throughput PCR
Amplification of DNA fragments up to 8 kb
Generation of PCR products for TA cloning
DNA labeling
Storage
Protected from light and avoid multiple freeze/thaw cycles 4°C for 6 months -20°C for 24 months
Document
The ExcelTaq™ 5X Fluorescent PCR Master Mix is a ready-to-use mixture for amplifying targeted DNA fragments. It is designed to serve as a master mix for virtually all PCR applications. This product is supplied as a 5X concentrated mixture containing all the essential ingredients for PCR with the exception of templates and primers. In addition, the mixture contains electrophoresis tracking dye (Bromophenol blue) and a safer fluorescent DNA staining dye, which enables the user to track the electrophoresis process in real time as well as eliminates the need for the staining process. The resultant PCR reaction mixture is sufficiently dense enough to be loaded directly into a TAE or TBE buffered gel for electrophoresis.
This product provides fast and easy methods for purification of total DNA for reliable PCR and Southern Blotting. Total DNA (e.g., genomic, viral, mitochondrial) can be purified from tissue and culture cells.
Details
Specifications
Features
Specifications
Main Functions
Isolation total DNA from 2ml blood and 200mg tissue using Midi column
Applications
PCR, southern bolt and virus detection, etc
Purification method
Midi spin column
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Tissue, cell, blood, saliva, swab, blood spot, semen and other clinical samples
Sample amount
0.2-2 ml
Elution volume
≥300μl
Time per run
≤80 minutes
Liquid carrying volume per column
4ml
Binding yield of column
1mg
Principles
This product is based on silica column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption column. Nucleic acid is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, Nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).
Advantages
High quality DNA – meet a variety of downstream applications, including PCR, qPCR, enzyme digestion, hybridization, etc.
Fast – without separation of leukocytes, organic extraction or ethanol precipitation
Simple – all nucleic acids can be obtained by direct digestion
Wide applicability – handle a variety of liquid samples
Kit Contents
Contents
D311302
D311303
Purification Times
20
100
HiPure gDNA Midi Columns
20
100
15ml Collection Tubes
40
200
Buffer ATL
50 ml
250 ml
Buffer AL
50 ml
250 ml
Buffer GW1*
22 ml
110 ml
Buffer GW2*
12 ml
50 ml
RNase A
20 mg
90 mg
Proteinase K
100 mg
440 mg
Protease Dissolve Buffer
10 ml
30 ml
Buffer AE
20 ml
120 ml
Storage and Stability
Proteinase K, RNase A should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions.
*Note：Leukocyte protocol can be used when large volume whole blood samples need to be processed. Whole blood was treated with red blood cell lysate, and white blood cells were obtained by centrifugation before extraction
Select the right purification kit to get impactful results：
This product provides fast and easy methods for purification of total DNA for reliable PCR and Southern Blotting. Total DNA (e.g., genomic, viral, mitochondrial) can be purified from tissue and culture cells.
Lyophilized format designed for room temperature shipping
Available in TaqMan format for analysis
Norgen’s Streptococcus agalactiae TaqMan PCR Lyophilized Kit is designed for the detection of Streptococcus agalactiae specific DNA in a real-time PCR based on the use of TaqMan® technology. The lyophilized format is designed to ship the kit at ambient temperature.
Norgen’s Streptococcus agalactiae TaqMan Lyophilized Probe/Primer and Control Set is designed for the detection of Streptococcus agalactiae specific DNA in a real-time PCR based on the use of TaqMan® technology. The lyophilized format is designed to ship the kit at ambient temperature.
All kit components should be stored at -20°C upon arrival.
Once reconstituted, repeated thawing and freezing (>2 times) 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 kit components can be stored for 2 years after the date of production without showing any reduction in performance.
