Komplet za ekstrakciju DNK gena krvi

1. Komponente kompleta reagensa

Tehnički podaci	50T	100T
Mačka. Ne.	SN0219	SN0220
Stupovi za ekstrakciju DNK (set)	50 (set)	100 (set)
Pufer reagensa b	20 ml	2 × 20 ml
Pufer reagensa c	30 ml	2 × 30ml
10×Red Blood Cell Lysis Buffer	60 ml	2 × 60ml
Pufer za pranje 1	15 ml	2 × 15 ml
Elucijski pufer	20 ml	20 ml
Proteinaza K	1ml	2x1ml
RNaseA	1ml	2x1ml
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 Blood Pročišćavanje DNK Reagent Kit offers a rapid and effective method for purifying DNA from blood and other bodily fluids. By lysing red blood cells with a red blood cell lysis buffer, DNA can be efficiently collected.

 

The Blood DNA Rapid Purification Kit can extract total DNA from whole blood (including blood, serum, plazma, and other bodily fluids) within 30 minuta. The entire purification process doesn’t require toxic reagents such as phenol-chloroform. The extracted DNA can be directly used for PCR, Južni blot, and other applications.

5. Eksperimentalni principi i postupci

6. Ekstrakcijski postupak

Prije početka eksperimenta:

A. Red Blood Cell Lysis Buffer 1x Preparation: Dilute the 10x Red Blood Cell Lysis Buffer to 1x volume using RNase-free water.

B. Reagent Buffer B and Reagent Buffer C may precipitate at low temperatures. It is recommended to heat at 65℃ for 5 minutes to dissolve the precipitate before use.

C. Prior to using Pufer za pranje 1, add the specified amount of anhydrous ethanol as indicated on the reagent bottle label, and mark a check on the label to indicate ethanol addition.

D. Buffer za eluciju je a 0.1X TE otopina with minimal EDTA. Ako EDTA može utjecati na naknadne eksperimente, it is suggested to substitute Elution Buffer with sterile deionized water.

1. Rukovanje uzorkom: (Collect 0.1-5 ml blood samples)

A. Dodati 2-3 times the volume of 1x Red Blood Cell Lysis Bufferto the blood or sample (Dilute the Red Blood Cell Lysis Buffer 10x to 1x before use), Temeljito pomiješajte, centrifuga na 12,000 RPM za 1 minuta, carefully remove the supernatant. The precipitate should theoretically be white or pale red. Add Reagent Buffer B to the precipitate (for 0.1-1ml blood samples, dodati 200μLPufer reagensa b; for 1-2ml blood samples, dodati 400μL Pufer reagensa b).

B. If handling samples from low-level organisms like poultry or birds with nucleated red blood cells, a smaller sample volume is needed. In such cases, omit the 1x Red Blood Cell Lysis Buffer and directly add 400μLof Reagent Buffer B and 20μL of RNaseA (10 mg/ml), Temeljito pomiješajte, and incubate at room temperature for 5 minuta.

2. Dodati 10μLRNaseA (10 mg/ml), 20μL Proteinaza K (10 mg/ml), Temeljito pomiješajte, digest at 65℃ for 10 minuta. Tijekom tog razdoblja, invert and mix 2-3 times until digestion is complete.

3. Dodati 200μLof Reagent Buffer C to the lysate, mix by pipetting, dodati 200μL of anhydrous ethanol, mix by pipetting.

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

5. Dodati 700μLof inhibition removal buffer, centrifuga na 12,000 RPM za 30 sekundi, odbaciti otpad.

6. Postavite stupac za pročišćavanje ekstrakcije DNK (set) in a new collection tube, dodati 500μLof Wash Buffer 1, centrifuga na 12,000 RPM za 30 sekundi, odbaciti otpad, and re-insert the DNA extraction purification column (set) into the collection tube for the next step.

(Bilješka: Ensure anhydrous ethanol has been added to Wash Buffer 1.)

7. Repeat step 6.

8. Postavite stupac za pročišćavanje ekstrakcije DNK (set) u novu cijev za centrifugu, uncover, incubate at 65℃ in a water bath for 2 minuta. Extend this step appropriately to evaporate ethanol as much as possible to prevent ethanol residue from affecting downstream experiments.

9. Suspend 50-100μLof Elution Buffer onto the membrane of the column, centrifuga na 12,000 RPM za 1 minuta, and collect the DNA.

(Bilješka: 1. Eluiranje DNK sa 50μL of Elution Buffer can increase DNA concentration but reduces total DNA yield; 2. The eluted DNA wash can be reapplied to the DNA extraction purification column. Centrifuga na 12,000 RPM za 1 minute again to increase DNA yield.