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)
Ampicillin and kanamycin selection markers
Storage
-20°C for 24 months
Other Products
Urine DNA Isolation Kits
Product Info
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Product Info
Overview
Rapid isolation of both small and large species of DNA from urine
Convenient spin column format
Effective removal of PCR inhibitors
Purified DNA is highly suited to sensitive downstream applications
Allows for the purification of viral DNA from urine
Both high molecular weight DNA (greater than 1 kb in size; mostly cell associated) and the smaller DNA (150 – 250 bp; derived from the circulation) is effectively isolated and purified using a rapid and convenient spin column protocol. This kit can be used to isolate DNA from a broad range of viruses in urine as well. Salts, metabolic wastes, proteins and other contaminants are removed to yield inhibitor-free DNA for use in sensitive applications. The DNA is of excellent quality for various downstream applications such as PCR, qPCR and DNA fingerprinting, methylation studies and more.
This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 50 μL to 1.75 mL of urine. Multiple samples can be processed in 30 minutes.
Urine DNA Isolation Kit (Slurry Format)
This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 3 mL to 25 mL. Multiple samples can be processed in 30 minutes. Multiple samples can be processed in 45 minutes.
Urine DNA Isolation Maxi Kit (Slurry Format)
This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 25 mL of urine up to 80 mL. Multiple samples can be processed in 45 minutes.
Background
DNA found in urine can be divided into 2 basic categories. The larger species, genomic-DNA (gDNA), is generally greater than 1 kb in size, and appears to be derived mainly from exfoliated cells. The second species is smaller, generally between 150 and 250 bp (apoptotic-DNA), and derives, at least in part, from the circulation. The second species is also considered as an RNA/DNA hybrid as reported by Halicka et al. (2000). Both types of DNA can be isolated reliably using this kit.
Storage Conditions and Product Stability All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm up Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
N-(t-butyl ester-PEG2)-N-bis(PEG2-propargyl) is a multi-functional PEG linker with two terminal propargyl groups and a t-butyl ester. The propargyl groups enables formation of triazole linkage with azide-bearing compounds or biomolecules in copper catalyzed Click Chemistry. The t-butyl protected carboxyl group can be deprotected under mild acidic conditions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
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N-(t-butyl ester-PEG2)-N-bis(PEG2-propargyl) is a multi-functional PEG linker with two terminal propargyl groups and a t-butyl ester. The propargyl groups enables formation of triazole linkage with azide-bearing compounds or biomolecules in copper catalyzed Click Chemistry. The t-butyl protected carboxyl group can be deprotected under mild acidic conditions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
For the determination of Escherichia coli in drinking water and its source water by multiple tube fermentation method.
Principle and Interpretation
MUG permits the rapid detection of Escherichia coli when the medium is observed for fluorescence using a UV light. MUG is hydrolyzed by the enzyme β-glucuronidase which is produced by E. coli to yield a fluorescent end product 4- methylumbelliferone. Trypticase provides the essential nutrients. Lactose is the fermentable carbohydrate. Sodium chloride maintains the osmotic equilibrium. The medium has a strong buffering system to control the pH in the presence of fermentative action. The bile salts inhibit gram-positive bacteria especially the Bacillus species and faecalStreptococci.
Formulation
Ingredients
/liter
Trypticase or tryptose
20 g
Bile salts No. 3
1.5 g
Lactose
5 g
K2HPO4
4 g
KH2PO4
1.5 g
NaCl
5 g
4-methylumbelliferyl-β-D-glucuronide (MUG)
50mg
pH6.9±0.2 at 25°C
Preparation
Weigh 37g of dry powder of this product, add 1L of distilled water or deionized water, stir, heat and boil until
completely dissolved, sterilize at 115℃ for 20min, cool to room temperature and set aside.
Quality Control
The following quality control strains were inoculated and cultured at 44.5℃±0.5℃ for 24h. The results are as follows:
Quality control strains
Growth
Escherichia coli ATCC25922
Blue-white fluorescence is produced under 366nm UV light
Salmonella typhimurium ATCC14028
No fluorescence under 366nm UV light
Enterococcus faecalis ATCC29212
Clear, no fluorescence
Storage and Shelf Life
2-30℃，Keep container tightly closed, avoid direct sunlight.
Use before expiry date on the label.
Precautions
1. When weighing the dehydrated medium, please wear masks to avoid causing respiratory system discomfort
2. Keep container tightly closed after using to prevent clumping.
Waste Disposal
Microbiological contamination was disposed by autoclaving at 121°C for 30 minutes.
Document
Intended Use For the determination of Escherichia coli in drinking water and its source water by multiple tube fermentation method. Principle and Interpretation MUG permits the rapid detec……
