Komplet za ekstrakciju životinjskog/staničnog genomskog DNA

1. Komponente kompleta reagensa

Tehnički podaci	50T	100T
Mačka. Ne.	SN0221	SN0222
Stupovi za ekstrakciju DNK (set)	50 (set)	100 (set)
Reagent Buffer IV	20 ml	2×20 ml
Reagent Buffer Cplus	30 ml	30 ml
Pufer za pranje 1	15 ml	2 × 15 ml
Elucijski pufer	20 ml	20 ml
Proteinaza K	1ml	2x1ml
RNaseA	1ml	1ml
Priručnik za upute	1	1

2. Skladištenje

Ovaj se komplet reagensa treba čuvati na sobnoj temperaturi (15-25℃) I u suhim uvjetima, s rokom trajanja 12 mjeseca. Stupovi za pročišćavanje DNK mogu se pohraniti za 1 godina u hladnom i suhom okruženju. Proteinase K and RNaza A contain preservatives, omogućavajući prijevoz na sobnoj temperaturi, Ali za dugoročno skladištenje, Treba ih držati na -20 ℃.

3. Upute za korištenje kompleta reagensa

3.1 Ovaj komplet reagensa namijenjen je istraživanju molekularne biologije i ne smije se koristiti za dijagnozu ili liječenje bolesti.

3.2 Neke komponente u kompletu reagensa sadrže iritante. Preporučuju se zaštitne mjere poput nošenja zaštitne odjeće i naočala.

3.3 Tijekom upotrebe ovog kompleta reagensa, brza centrifuga, vodena kupelji (metalna kupka), vrtlog miksera, bezvodni etanol, sterilna deionizirana voda, i EP epruvete treba pripremiti korisnik.

4. Uvod u komplet reagensa

The Animal/Tissue Pročišćavanje DNK Kit offers a rapid and effective solution for animal tissue and cell culture DNA purification, widely applicable across animal tissues and cell cultures.

This DNA rapid purification kit swiftly extracts total DNA from animals and cells, yielding DNA directly usable for PCR, Južni blot, and similar applications. The purification process does not require toxic agents like phenol-chloroform, making this DNA purification kit highly suitable for various other samples.

5. Eksperimentalni principi i postupci

6. Ekstrakcijski postupak

Prije početka eksperimenta:

A. Reagent Buffer IV tends to precipitate under low-temperature conditions. It is recommended to heat at 65℃ for 5 minuta. After the precipitation dissolves, Može se normalno koristiti.

B. Prior to use, Dodajte navedenu količinu bezvodnog etanola u PranjeBuffer 1 as indicated on the reagent bottle label, and mark a check on the label to indicate the addition of anhydrous ethanol.

C. Buffer za eluciju je a 0.1X TE otopina with minimal EDTA content. Ako EDTA može utjecati na naknadne eksperimente, it is recommended to replace the Elution Buffer with sterile deionized water.

1. Rukovanje uzorkom:

A. Take cell culture, centrifuga na 12,000 RPM za 1 minute to collect cells, and aspirate the supernatant as much as possible. Add 250μl of Reagent Buffer IV and 10μl of RNaseA (10 mg/ml) to the collected cells, thoroughly suspend, and incubate at 65°C for 10 minuta.

B. Grind tissue not exceeding 25 mg into a fine powder using liquid nitrogen. Add 200μl of Reagent Buffer IV, 20μl proteinaze k (10 mg/ml), and 10μl of RNaseA (10 mg/ml) to the powder, shake well, and incubate at 65°C for 30 minuta, shaking the mixture four times during this period.

2. Dodati 200μl of Reagent Buffer Cplus to the lysate and mix well. Ako se pojavi bijeli talog, it can be left undisturbed; To neće utjecati na naknadne eksperimente nakon što talog nestane.

3. Dodati 200μl of anhydrous ethanol and mix thoroughly. Some precipitation might occur, but it won’t affect subsequent experiments.

4. Apply the obtained liquid to the DNA extraction purification column (sleeve) (approximately 650~700μl each time), let it sit at room temperature for 2 minuta, centrifuga na 12,000 RPM za 30 sekundi, discard the collected waste, and re-insert the collection tube into the purification column for the next step.

5. Postavite stupac za pročišćavanje ekstrakcije DNK (sleeve) into a new collection sleeve, dodati 700μl of PranjeBuffer 1, centrifuga na 12,000 RPM za 30 sekundi, odbaciti otpad, and place the DNA extraction purification column (sleeve) back into the sleeve for the next step.

(Bilješka: Confirm the addition of anhydrous ethanol to Rinse Buffer 1.)

6. Dodati 500μl of PranjeBuffer 1 na stupac za pročišćavanje DNK (sleeve), centrifuga na 12,000 RPM za 30 sekundi, extend centrifugation time appropriately to ensure the membrane is drier.

(Bilješka: Ethanol has a significant impact on subsequent experiments; therefore, membrane dryness is crucial. After centrifugation, ensure no ethanol remains before elution. Discard the waste and collection tubes afterward. After rinsing with Wash Buffer 1, the membrane on the DNA purification column should have a faint color. Carefully remove the DNA purification column after centrifugation, ensuring it doesn’t touch the collection tube to prevent ethanol contamination.)

7. Place the DNA purification column in a new centrifuge tube, add 100μl of Elution Buffer directly onto the membrane, centrifuga na 12,000 RPM za 1 minuta, and collect the DNA.

(Bilješka: Eluting DNA with 50μl of Elution Buffer can increase DNA concentration but decreases total DNA yield.)