MagBind Bacterial Total RNA Extraction Kit (Prepackaged 48T) servicebio G3699-48T
$50.00
In stock
SKU
G3699-48T
Product Information
| Product Name | Cat.No. |
| Spec. | MagBind Bacterial Total RNA Extraction Kit (Prepackaged 48T) |
| G3699-48T | 48T |
Description/Introduction
This product is 48 prepackaged plates, which uses Servicebio Y-16, Y-48 automatic nucleic acids extractor and self-developed superparamagnetic magnetic beads to have strong reversible adsorption of nucleic acids under certain buffer conditions. It is designed for total RNA extraction of 48 samples from a variety of Gram-negative bacteria and most Gram-positive bacteria, and the entire process can be completed in less than 1 hour, replacing the current time-consuming and cumbersome isopropanol precipitation steps and a large number of rinsing steps. DNase contains visual dyes that have a sample tracing effect, preventing DNase from being missed and resulting in genomic DNA residue in the extracted RNA. The extracted RNA has good integrity, high purity, and large yield, and can be directly used in various molecular biology experiments such as RT-PCR, Real-Time PCR, Chip analysis, Northern Blot, Dot Blot, PolyA screening, In vitro translation, RNase protection analysis and molecular cloning.
Storage and Handling Conditions
Lysozyme (50 mg/mL), DTT solution and DNase are shipped with wet ice and stored at -20°C. Other reagents are shipped and stored at room temperature; valid for up to 12 months.
Product Contents
| Component Number | Component |
| G3699-48T | G3699-1 |
| Lysozyme (50 mg/mL) | 500 μL |
| G3699-2 | Buffer GRL |
| 15 mL | G3699-3 |
| DTT Solution | 600 μL |
| G3699-4 | DNase |
| 500 µL | G3699-5 |
| 96-well prepackaged plate | 3 |
| G3699-6 | 8-hole magnetic rod sleeve |
| 6 | Manual |
| 1 copy |
Before starting (please read carefully).
1. Check if there is any leakage of the prepackaged plates and discrepancy in the instruction manual. If such a phenomenon occurs, do not use it.
2. Before use, observe whether the magnetic beads or reagents are stuck to the sealing film or the hole wall of the reagent prepackaged plates. If this phenomenon occurs, you can mix them up and down and then centrifuge briefly or gently tap the bottom of the prepackaged plates on the table until the droplets fall off before use.
3. Please prepare 10 mM Tris-HCl (pH 8.0) for resuspension of collected bacterial cells and dilution of lysozyme.
4. If extracting Gram positive bacterial RNA, please prepare a 37℃ water bath or heating block in advance.
5. If a precipitate has formed in Buffer GRL, warm it at 37°C until the precipitate has fully dissolved. Use it after cooling down to room temperature.
6. Before operation, add appropriate volume of DTT Solution into Buffer GRL to a final concentration of 4%. Add 40 µL DTT Solution per 1 mL of Buffer GRL. It should be prepared immediately before use. This mixture can be stored at 4℃ for one month.
7. Add 10 µL of DNase (blue) to the liquid (yellow) in the 4th and 10th columns of the prepackaged plates, and the liquid will be green after adding DNase.
Assay Protocol / Procedures
1. Take 1-3 mL of bacterial solution (the total amount of bacteria does not exceed 1×109) and centrifuge at 12,000 rpm (~13,400×g) at 4°C for 2 min to collect the bacteria.
Note: The supernatant must be removed clean, otherwise it will affect the digestion of bacterial cell wall.
Note: Bacteria should not be excessive, otherwise it will cause problems such as yield reduction, genome residue and protein contamination. When OD600=1, it is recommended to take the amount of bacterial culture of 1 mL.
2. Suspend the bacterial body weight in 100 μL of 10 mM Tris HCl (pH 8.0) containing Lysozyme. The Lysozyme concentration and incubation conditions are shown in the table below:
| Bacterial categories | Lysozyme concentration |
| Incubation conditions | Incubating time |
| Gram-negative bacteria(G-) | 0.4 mg/mL |
| Room temperature | 3-5 min |
| Gram-positive bacteria(G+) | 5 mg/mL |
| 37℃ | 5-10 min |
3. Add 300 μL of Buffer GRL to the incubated bacterial suspension (confirm DTT solution was added before use), and mix thoroughly with vortex oscillation;
4. Preparation of prepackaged plates: Take out the prepackaged plate, centrifuge it at 500 rpm for 1min, or gently tap the bottom of the prepackaged plate on the table until the liquid drops off, and then carefully tear off the aluminum foil sealing film;
5. Add 10 µL of DNase (blue) to the liquid (yellow) in the 4th and 10th columns of the prepackaged plates, and the liquid will be green after adding DNase;
6. Transfer the mixture in step 3 to the 1th and 7th columns of the prepackaged plates;
7. Place the prepackaged plates on the corresponding position of the nucleic acid extractor;
8. Parameter settings of Servicebio Y-16 and Y-48 automatic nucleic acid extractor:
| Step | First |
| Second | Third |
| Fourth | Fifth |
| Sixth | Seventh |
| Eighth | Ninth |
| Station | 3 |
| 1 | 2 |
| 3 | 4 |
| 3 | 5 |
| 6 | 5 |
| Wait Time | 00:00:00 |
| 00:00:00 | 00:00:00 |
| 00:00:00 | 00:03:00 |
| 00:00:00 | 00:00:00 |
| 00:05:00 | 00:00:00 |
| Mix Modea | 2 |
| 2 | 2 |
| 2 | 2 |
| 2 | 2 |
| 2 | 2 |
| Mix Time | 00:00:10 |
| 00:05:00 | 00:01:00 |
| 00:01:00 | 00:10:00 |
| 00:01:00 | 00:01:00 |
| 00:05:00 | 00:00:10 |
| Pause | Ⓧ |
| Ⓧ | Ⓧ |
| Ⓧ | Ⓧ |
| Ⓧ | Ⓧ |
| Ⓧ | Ⓧ |
| Mag Timea | 00:00:30 |
| 00:00:30 | 00:00:30 |
| 00:00:30 | 00:00:30 |
| 00:00:30 | 00:00:30 |
| 00:01:00 | 00:00:00 |
| Volume (µL) | 600 |
| 500 | 500 |
| 600 | 100 |
| 600 | 600 |
| 80 | 600 |
| Temperature (℃) | --- |
| 25.0℃ | --- |
| --- | --- |
| --- | --- |
| --- | --- |
a: If you do not use the Servicebio Y-16 and Y-48 automatic nucleic acid extractor, you can adjust the magnetic attracting time and mixing speed parameters according to other instrument instructions suitable for this product.
9. Install the magnetic sleeve, run the program;
10. After the automated extraction steps, transfer the nucleic acid solution from the 6th and 12th columns to a new Nuclease-free centrifuge tube carefully and store at-80℃ for long-term storage.
Note
1. Please read this product manual carefully before operating.
2. Please do not invert the prepackaged plates.
3. Do not dry magnetic beads for a long time, so as not to affect the efficiency of RNA elution.
4. During the experiment, experimental clothing, disposable gloves, and masks should be worn to avoid speaking. Please ensure that the equipment used for RNA extraction and the water or reagents used for preparing the solution are Nuclease free products.
5. The degree of enzymatic hydrolysis of different bacterial cell walls varies, and Gram-positive bacterial cell walls are generally more difficult to hydrolyze. Customers can adjust the concentration of lysozyme and incubation time according to the type of bacteria.
6. RNA extraction specialized experimental platforms and electrophoresis equipment should be used.
For Research Use Only!
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