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The Heat&Run® gDNA removal kit removes contaminating gDNA from RNA prior to RT-qPCR.

The kit is based on the recombinant heat labile dsDNase, which is irreversibly inactivated at low temperatures (5 minutes at 58ºC or 15 minutes at 55ºC).

Reverse Transcription can be performed in the same tube, thereby minimizing pipetting steps and reducing hands-on time.

Protocol

The kit contains HL-dsDNase which efficiently removes gDNA from RNA preps before running RT-qPCR.

• gDNA is removed, leaving RNA ready for reverse transcription in the same tube (Figure 1).
• Heat-labile dsDNase can easily be inactivated.
• This procedure minimizes pipetting steps and reduces hands-on time.
• The Heat&Run kit is especially well suited for high throughput experiments.

The high activity of the HL-dsDNase at low temperature and its heat-lability makes it ideal to use in a combination with a heat activated RT enzyme. The contaminating genomic DNA is cleaved at ambient temperature and increasing the temperature over 50°C will inactivate the DNase and start the Reverse Transcriptase.

Do you require gDNA removal in applications other than RT-qPCR? Contact our support team for assistance in implementing dsDNase treatment in your workflow.

Kit Contents

• 10X reaction Buffer
• HL-dsDNase (50 or 250 reactions)

The Heat&Run® gDNA removal kit removes contaminating gDNA from RNA prior to RT-qPCR.

The kit is based on the recombinant heat labile dsDNase, which is irreversibly inactivated at low temperatures (5 minutes at 58ºC or 15 minutes at 55ºC).

Reverse Transcription can be performed in the same tube, thereby minimizing pipetting steps and reducing hands-on time.

Introduction

The MagPure Plasmid purification system uses the paramagnetic bead technology for high-throughput preparation of high-copy or low-copy plasmid DNA from E. coli cells. This kit also can be used with fosmid and BAC vector-based constructs. The system uses alkaline lysis followed by a MagPure purification to differentially bind plasmid DNA to paramagnetic beads. While the DNA is bound to the beads, contaminants can be rinsed away using a simple washing procedure. Because MagPure uses magnetic separation technology, the protocol does not require vacuum filtration. This makes kit extremely amenable to automation. Plasmid DNA purified with this system is most commonly used in Sanger Sequencing and PCR amplification.

Details

Specifications

Features	Specifications
Main Functions	Isolation up to 15μg endotoxin free plasmid DNA from 1-5ml bacterial culture
Applications	Enzyme digestion, sequencing, PCR and labeling,  etc.
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	Conventional plasmid, plasmid≤30KB
Sample amount	1-5ml
Elution volume	≥50μl
Time per run	≤80 minutes

Principle

This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Lysozyme. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption.The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by Elution Buffer.

Advantages

• High purity – purified plasmid can be directly used in sequencing, enzyme digestion and PCR, etc.
• Fast – it takes only 80 minutes to complete the isolation
• High yield – up to 15μg plasmid can be binded in one column
• Economic – excellent cost effectiveness performance

Kit Contents

Contents	P181402	P181403	P181404
Purification Times	100 Preps	500 Preps	5000 Preps
RNase A	10 mg	30 mg	2 x 160 mg
Buffer P1	30 ml	150 ml	2 x 800 ml
Buffer P2	30 ml	150 ml	2 x 800 ml
Buffer LEN3	20 ml	80 ml	800 ml
Buffer LN4	90 ml	400 ml	4 x 980 ml
MagPure Particles	3.5 ml	17 ml	3 x 60 ml

Storage and Stability

RNase A and MagPure Particles should be stored at 2–8°C upon arrival. However, short-termstorage (up to 12 weeks) at room temperature (15–25°C) does not affect its performance. Theremaining kit components can be stored dry at room temperature (15–25°C) and are stable for atleast 18 months under these conditions.The entire kit can be stored at 2–8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature whenused. If any precipitates form in the buffers,warm at 37℃ to dissolve. After addition of RNase A, Buffer P1 is stable for 6 months when stored at

The MagPure Plasmid purification system uses the paramagnetic bead technology for high-throughput preparation of high-copy or low-copy plasmid DNA from E. coli cells. This kit also can be used with fosmid and BAC vector-based constructs. The system uses alkaline lysis followed by a MagPure purification to differentially bind plasmid DNA to paramagnetic beads. While the DNA is bound to the beads, contaminants can be rinsed away using a simple washing procedure. Because MagPure uses magnetic separation technology, the protocol does not require vacuum filtration. This makes kit extremely amenable to automation. Plasmid DNA purified with this system is most commonly used in Sanger Sequencing and PCR amplification.

Introduction

Usages:

For the rapid detection and enumeration of Escherichia coli.

Principle:

Peptone and yeast extract powder provides carbon and nitrogen sources and trace elements; sodium chloride maintains osmotic equilibrium; agar as medium coagulant; dodecyl sulfate inhibit Gram-positive bacteria; chromogenic substrate and large intestine coli β- glucuronidase enzyme specific reaction, hydrolysis of the substrate, the release of the color groups produce green colonies on the light yellow plate.

Formulation (per liter):

Peptone :15.0g

Yeast extract powder: 3.0g

Sodium chloride: 5.0g

Sodium lauryl sulfate:0.1g

Agar: 12.0g

Chromogenic substrate 6.5g

Final pH 7.0 ± 0.2

How to use:

1. Weigh 41.6g of the product, adding 1.0L distilled or deionized water, heated to boiling stirring until completely dissolved, dispensing into flask, 115 autoclaved for 10minutes.

2. Take 25.0g or 25.0ml of sample with sterile procedures , added  to the flask containing 225.0mL of sterile phosphate buffered saline (or saline) , shaken thoroughly homogenized with a homogenizer or a 1:10 dilution of 1min solution, diluted 1:10 and then continue to select the appropriate serial dilutions of three, the two plates inoculated with each dilution, poured dissolved by heating and cooled to about 45 medium.

3, observe the results.

Quality Control

This product appears light yellow after the pouring on plate, these strains were inoculated after 36 ± 1 18 ~ 24h culture growth in the following table.

Bacteria name            Bacteria NO.       Growth Situation         Feature

Escherichia coli          ATCC25922          good            green colonies

Citrobacter              ATCC8090           good           colorless colonies

Salmonella typhimurium   CMCC50115         good            colorless colonies

Enterococcus faecalis     ATCC29212         suppressed                —–

Storage: Store in a dark, cool and dry place, tighten the caps immediately after use. Storage period of two years.

1000mL