EasyPure вирусная ДНК/РНК экстракция нуклеиновой кислоты реагент от TransGen Biotech

Описание и отзывы

Характеристики

EasyPure^® Viral DNA/RNA Kit

EasyPure^® Viral DNA/RNA Kit utilizes a unique lysis buffer to lyse virus and release DNA / RNA. The released DNA / RNA is effectively purified after specifically binding to a silica-based spin column. It is suitable for isolating viral DNA/RNA from up to 200 μl of plasma, serum, whole blood, tissue homogenate, cell-free body fluid, nasopharyngeal or oropharyngeal aspirate/wash, bronchoalveolar lavage fluid (BALF), tracheal aspirate, sputum, nasopharyngeal or oropharyngeal swab and animal cell culture supernatant. The isolated DNA/RNA with high purity can be applied in PCR, RT-PCR, qPCR, qRT-PCR, etc.

Kit Contents

Sample requirement

• Storage: at 2-8^oC for 4 hours; at -20^oC or -80^oC for long term storage.

• Avoid repeated freezing and thawing for plasm and serum (no more than once)

Procedure

Before starting, add 48 ml of 96%-100% ethanol to the 12 ml WB5, mix thoroughly and mark it. Prepare BB5 with carrier RNA according to the schedule given in next page. All centrifugation steps were carried out at room temperature.

1. Add 20 μl Proteinase K to a sterile 1.5 ml microcentrifuge tube.

2. Add 200 μl sample to tube. (Note: If the sample volume is less than 200 μl , please add PBS or 0.9% NaCl to 200 μl).

3. Add 200 μl BB5 with carrier RNA, mix by vortexing for 15 seconds.

4. Incubate at 56 ^oC for 15 minutes.

5. Add 250 μl of 96%-100% ethanol to the sample (there may appear floculation). Mix by vortexing for 15 seconds and incubate at room temperature (15-25^oC) for 15 minutes.

6. Add all lysate to a spin column and centrifuge at 12,000×g for 1 minute, discard flow through.

7. Add 500 μl WB5 (check to be sure you have added ethanol). Centrifuge at 12,000×g for 1 minute, discard flow through.

8. Repeat step 7.

9. Centrifuge at 12000×g for 1 minute to eliminate residual ethanol and air-dry the membrane completely. 10. Place the spin column into a clean RNase-free 1.5 ml microcentrifuge tube. Add 20-50 μl of RNase-free Water to the center of the column, and incubate at room temperature for 1 minute.

11. Centrifuge at 12000×g for 1 minute. 1

2. Store the eluted DNA (at -20^oC) or RNA (at -80^oC).

Addition of carrier RNA to BB5

Calculate the volume of BB5 with carrier RNA needed by selecting the number of samples to be simultaneously processed from Table 1.

Gently mix by inverting the tube 10 times. To avoid foaming, do not vortex.

For larger numbers of samples, volumes can be calculated using the following sample calculation:

N × 0.21 ml = A ml

A ml × 28 μl/ml = B μl

where: N = number of samples to be processed simultaneously

A = calculated volume of BB5

B = volume of carrier RNA to add to BB5

Table 1. Volumes of BB5 and carrier RNA required for different number of samples.

Notes

• All the centrifugation steps are carried out at room temperature.

• Please check to ensure you have added 96%-100% ethanol into WB5 before use.

• Aliquot the Carrier RNA into RNase-free microcentrifuge tubes and store at -20^oC. Avoid repeated freezing and thawing (not more than three times) for Carrier RNA.