[CK1000] Champion™ E. coli Transformation Kit, 200 Rxn
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Champion™ E. coli Transformation Kit provides an easy method for rapid preparation of chemically competent cells with high transformation efficiency from fresh culture, overnight culture, or even directly from bacterial colonies on the plate. The competent cell preparation method eliminates the requirement of time-wasting wash step. In addition, preparation of competent cells from overnight culture or directly from bacterial colonies provides flexibility to cloning experiment. The resultant competent cells can be immediately used or stored at -70°C for one year. This kit includes a specialized SMO-Broth™ medium and a unique Champion™ CC Buffer for culturing and preparing competent cells efficiently. Following the simple and quick competent cell preparation protocol from fresh culture, the transformation efficiency is typically ranged from 108–109 cfu/μg transformants/μg of pUC19 plasmid DNA, but varies depending on the E. coli strains. The resultant competent cells can be further transformed using time-saving transformation protocol, eliminating the requirement of heat-shock and recovery steps.
Detail
Description
Champion™ E. coli Transformation Kit provides an easy method for rapid preparation of chemically competent cells with high transformation efficiency from fresh culture, overnight culture, or even directly from bacterial colonies on the plate. The competent cell preparation method eliminates the requirement of time-wasting wash step. In addition, preparation of competent cells from overnight culture or directly from bacterial colonies provides flexibility to cloning experiment. The resultant competent cells can be immediately used or stored at -70°C for one year. This kit includes a specialized SMO-Broth™ medium and a unique Champion™ CC Buffer for culturing and preparing competent cells efficiently. Following the simple and quick competent cell preparation protocol from fresh culture, the transformation efficiency is typically ranged from 108–109 cfu/μg transformants/μg of pUC19 plasmid DNA, but varies depending on the E. coli strains. The resultant competent cells can be further transformed using time-saving transformation protocol, eliminating the requirement of heat-shock and recovery steps.
Features
Flexible– fresh culture, overnight culture, 4°C stored liquid culture or even colonies on agar plate can be used for transformation.
Fast and Easy– only few steps for preparation; suitable for time-saving transformation
High efficiency– up to 109 cfu/μg
Personalization– suitable for most E. coli strains
Kit Contents
Component
Volume
Champion™ CC Buffer
20 ml
SMO-Broth™
100 ml x 2
pUC19 Control Plasmid (10-4 μg/μl)
5 µl
Instruction Manual
1
Champion™ Competent Cell Preparation Card
1
Storage
4°C for 12 months
Other Products
AAV Purification Kits
Product Info
Document
Product Info
Overview
AAV Purification from any input – cell fraction or media fraction
High AAV recovery, up to 90%
No specialized equipment needed
Purification from a variety of AAV serotypes (including AAV6 and AAV9)
Yields highly active AAV for in vivo and in vitro experiments
Purification is based on spin column chromatography that uses Norgen’s resin separation matrix
Recombinant adeno-associated virus (AAV) vectors are highly promising tools for both in vitro and in vivo gene transfer. Norgen’s AAV Purification Kits provide fast and simple procedures for concentrating and purifying AAV vectors from cell lysate and cell culture media. Purification is based on precipitation onto Norgen Biotek’s proprietary resin. Contaminating cellular debris is largely removed from the sample via a centrifugation step, while contaminating DNA and RNA is reduced using enzymatic digestion. AAV vector purified in this manner is highly active for use in in vitro and in vivo transduction experiments.
AAV Purification Kit
Norgen’s AAV Purification Kit contains sufficient materials for 15 preparations (33.5 mL per prep of supernatant (SN) or a total of 500 mL of supernatant input). Approximately 1 mL of cell pellet can be purified per prep, up to a maximum of 15 mL of cell pellet in total for the entire kit. Up to 33X sample concentration.
AAV Purification Mini Kit
Each spin column is able to concentrate and purify AAV from 0.5-8 mL of cell pellet, cell culture media, or cells and culture media mixed together. Up to 50X sample concentration. AAV vector purified in this manner is highly active for use in in vitro transduction experiments, and is eluted into a small volume (200 µL). Preparation time for 4 samples is 1.5 hours, with 45 minutes of hands-on time.
AAV Purification Midi Kit
Each spin column is able to concentrate and purify AAV from 8 mL up to 45 mL of input consisting of cell pellet, cell culture media, or cells and culture media mixed together. Up to 50X sample concentration. AAV vector purified in this manner is highly active for use in in vitro transduction experiments, and is eluted into a small volume (1 mL). The kit may be used to purify up to 8 x 25 mL or 4 x 45 mL of samples using the included columns. Preparation time for 4 samples is approximately 2 to 2.5 hours, with 1.5 hours of hands on time.
AAV Purification Maxi Kit (Slurry Format)
Each spin column is able to concentrate and purify AAV from 45 mL to 90 mL of input consisting of cell pellet, cell culture media, or cells and culture media mixed together. Up to 200X sample concentration. AAV vector purified in this manner is highly active for use in in vitro transduction experiments, and is eluted into a small volume (1-10 mL) using the optional concentration step. The kit may be used to purify up to 1 x 900 mL samples or 10 x 45-90 mL samples using the included columns. Preparation time for 1 x 900 mL sample is approximately 2.5 to 3.5 hours, with an optional concentration step requiring an additional 30 min.
At least 1 x 1010 AAV particles as determined by qPCR
AAV Vector Serotype
Any
Average Recovery
> 80%
Input Type
Cells, media, or mixed
Input Volume
8 mL – 45 mL
Minimum Elution Volume
1 mL
Time to Complete 4 Purifications
2 – 2.5 hours
Storage Conditions and Product Stability DNAse I and RNAse A should be stored at -20°C upon arrival. Elution Buffer O should be stored tightly capped at 4°C upon arrival. All other solutions should be kept tightly sealed and stored at room temperature. Once opened, the solutions should be stored at 4°C. This kit is stable for 1 year after the date of shipment.
Component
Cat. 66100 (15 preps)
Cat. 63200 (20 preps)
Cat. 63300 (4-8 preps)
Cat. 63250 (1-10 preps)
Lysis Buffer S
5.5 mL
5.5 mL
5.5 mL
20 mL
DNAse I
–
2 x 25 uL
2 x 25 uL
210 μL
RNAse A
–
60 μL
60 μL
240 μL
HL-SAN Nuclease
102 μL
–
–
–
Binding Buffer A
20 mL
4 mL
4 mL
2 x 8 mL
Purification Solution C
60 mL
–
–
–
Purification Solution D
130 mL
–
–
–
Wash Solution C
2 x 130 mL
60 mL
60 mL
3 x 60 mL
Slurry E
12.5 mL
–
–
2 x 14.5 mL
Elution Buffer O
66 mL
8.5 mL
8.5 mL
66 mL
Protein Neutralizer
4 mL
4 mL
4 mL
4 mL
Spin Columns
–
20
–
–
Mini Spin Columns
–
20
–
–
Midi Spin Columns (grey contents) with Collection Tubes
–
–
8
10
Midi Spin Columns (white contents) with Collection Tubes
–
–
8
–
Maxi Spin Columns (grey contents) with Collection Tubes
–
–
–
10
Maxi Spin Columns (white contents) with Collection Tubes
The Kit provides fast purification of high-quality RNA from paxigene Blood RNA Tube using silica-membrane spin columns. RNA purified using the HiPure System is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.
The Kit is for the purification of total RNA from 2.5 ml human whole blood collected in a PAXgene Blood RNA Tube. Purification begins with a centrifugation step to pellet nucleic acids in the PAXgene Blood RNA Tube. The pellet is washed and resuspended, and incubated in optimized buffers together with proteinase K. DNA wash The lysate is passed through a DNA Mini column. Ethanolis added to adjust binding conditions, and the lysate is applied to a column.RNA is selectively bound to the silica membrane as contaminants pass through. Remaining contaminants are removed in several efficient wash steps. Between the first and second wash steps, the membrane is treated with DNase I to remove trace amounts of bound DNA. After the wash steps, RNA is eluted in elution buffer and heat-denatured.
Advantages
High quality – high purity total RNA can be directly used in various sensitive downstream applications
Fast – isolation of several samples can be completed in 40 minutes by using column purification method
Safety – no phenol chloroform extraction required
Sensitive – direct lysis of blood, plasma and other samples without separation of leukocytes
Kit Contents
Contents
R416802
R416803
Purification Times
10 Preps
50 Preps
HiPure RNA Mini Columns I
10
50
gDNA Filter Mini Columns
10
50
2ml Collection Tubes
30
150
RNase Free Water
60 ml
250 ml
Buffer MBR1
10 ml
30 ml
Buffer MBR2
5 ml
15 ml
Buffer RW1
15 ml
60 ml
Buffer RW2*
6 ml
20 ml
Proteinase K
12 mg
50 mg
Protease Dissolve Buffer
1.8 ml
5 ml
DNase I
120 µl
600 µl
DNase Buffer
6 ml
30 ml
Storage and Stability
Proteinase K should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, Proteinase K up to 8 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.
Experiment Data
Document
The Kit provides fast purification of high-quality RNA from paxigene Blood RNA Tube using silica-membrane spin columns. RNA purified using the HiPure System is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.
The SMO-HiFi™ DNA Polymerase is a new genetically modified, recombinant DNA polymerase with fidelity 70 times higher than Taq DNA polymerase during amplification, as well as very high elongation rate. Being highly thermostable, SMO-HiFi™ DNA Polymerase can remain viable even after being subjected to boiling for 2 minutes. The SMO-HiFi™ DNA Polymerase is also designed to operate in much lower Mg2+ concentration as compared to other DNA polymerase products.
Features
5’→3′ DNA polymerase activity
3’→5′ exonuclease (proofreading) activity
High reaction rate (up to 1 kb/10 seconds)
High fidelity, 70 times higher than Taq DNA polymerase
Blunt end amplicons
Thermo-stable: half-life is more than 10 hrs at 95°C
Storage
[TF1000] SMO-HiFi™ DNA Polymerase
-20°C for 24 months
Document
The SMO-HiFi™ DNA Polymerase is a new genetically modified, recombinant DNA polymerase with fidelity 70 times higher than Taq DNA polymerase during amplification, as well as very high elongation rate. Being highly thermostable, SMO-HiFi™ DNA Polymerase can remain viable even after being subjected to boiling for 2 minutes. The SMO-HiFi™ DNA Polymerase is also designed to operate in much lower Mg2+ concentration as compared to other DNA polymerase products.
