Product Introduction

Product Name	Cat.No.	Spec.
Blood Direct PCR Kit	G3312-01	40 rxns
Blood Direct PCR Kit	G3312-05	200 rxns

Description/Introduction

The kit can directly amplify whole blood samples without DNA extraction or sample preparation. It is suitable for fresh blood, blood stored at 4°C or frozen containing conventional anticoagulants such as EDTA, heparin and citrate, and dried blood stains on Whatman903^® and FTA^® cards, genomic and exogenous target DNA can be effectively amplified.

After a series of optimization, the 2×Blood Direct PCR Master Mix provided by this kit, combined with the provided Blood Direct PCR Enhancer, showed a very high anti-inhibitory effect on blood direct expansion, and the amplification length could reach 5 kb. In addition, with the addition of Loading Dye in this product, the PCR product can be directly used for agarose gel electrophoresis. The product can be widely used in the identification of high-throughput blood samples.

Storage and Handling Conditions

2×Blood Direct PCR Master Mix should be shipped with wet ice, and stored at -20°C; Other reagents are shipped with wet ice, and stored at 2~8°C; valid for up to 12 months.

Product Contents

Component Number	Component	G3312-01	G3312-05
G3312-1	2×Blood Direct PCR Master Mix	1 mL	5×1 mL
G3312-2	Blood Direct PCR Enhancer	500 μL	5×500 μL
G3312-3	Nuclease-free Water	1 mL	5 mL
Manual		One copy

Assay Protocol / Procedures

1. Recommended PCR reaction system

Component	50 μL rxn	Final Concentration
2×Plant Direct PCR Master Mix	25 μL	1×
Blood Direct PCR Enhancer	10 μL	1×
Forward Primer (10 μM)^a	2 μL	0.4 μM
Reverse Primer (10 μM)^a	2 μL	0.4 μM
Template^b	Variable (≤10 μL)
Nuclease-free Water	Add to 50 μL

a. The final concentration of primers ranges from 0.2 μM to 1.0 μM, and the recommended concentration of primers is 0.4 μM. Too few primers may lead to low amplification yield or no amplification, and excessive primers may lead to non-specific amplification.

b. The amount of blood can be added between 1~20%, and 5% is recommended. Dry blood stains on Whatman903^®and FTA^® commercial cards can be cut to 1~2 mm² size using a punch or scissors. Note: in order to achieve the best PCR effect, use the pipette to slowly add the blood to the reaction system and precipitate it to the bottom of the reaction tube. After adding the blood sample, the reaction system cannot be mixed. The blood card can be added to the reaction tube in advance, and then the rest of the reaction system can be added to ensure that it is immersed in the reaction system.

2. Recommended PCR conditions:

Step	Temperature	Time	cycles
Initial Denaturation^a	98°C	10 min	1
Denaturation	98°C	10 sec	35
Annealing	50~65°C	20 sec
Extension^b	72°C	10~30 sec/kb
Final extension	72°C	5~10 min	1
Hold	4~16°C	Forever

a. The suitable annealing temperature can be selected according to the Tm value of the primer.

b. The extension time of shorter fragment (≤ 2kb) is 15 s/kb; the extension time of longer fragment (2~5 kb) is 30 s/kb.

Note

1. Please read the Product Manual carefully before use.

2. After amplification, centrifuge at 1,000×g or 4,000 rpm for 1~3 min, using the supernatant for electrophoresis.

3. For long-term storage of anticoagulant blood, blood clots should be avoided as far as possible.

4. In general, a good PCR effect can be achieved when the whole blood concentration is 5%. If not, the blood concentration can be adjusted between 1~20% according to the actual situation.

5. Birds or other nucleated red blood cells can reduce the amount of blood.

6. For the preparation and sub-packaging of the reaction solution, please use a new pipette tip to avoid cross contamination.

For Research Use Only!

Blood Direct PCR Kit