HCY™ Blood Direct PCR Kit

The HCY™ Blood Direct PCR Kit is a kit for direct PCR amplification of whole blood samples without the need for DNA purification or sample preparation. It can be used to directly amplify fresh, chilled (frozen) and commercially dried Whatman903® and FTA® blood stains containing conventional anticoagulants such as EDTA, heparin and citrate. The kits contain genetically engineered BloodDirect DNA Polymerase, which is highly resistant to PCR inhibitors in whole blood samples.

Uses
Direct PCR reaction of different types of whole blood samples.

Precautions
The recommended amount of blood template to be used is 10% of the total reaction volume, i.e. 5ul of whole blood is added to 50ul of reaction system;
Aspirate anticoagulated blood, blood clots should be avoided as much as possible;
The extension time is set at 30sec/kb, if it is less than 15sec, set 15sec can be used.
After the PCR reaction is completed, it is recommended that the reaction solution be centrifuged at 4000rpm (1000×g) for 1-3min to precipitate cellular debris, after which the supernatant is taken for downstream analysis.

Definition of activation
Using activated salmon sperm DNA as template/primer, the activity of 10 nmol of whole nucleotides ingested as acid insoluble material at 74°C for 30 min was defined as 1 activity unit (U).

CAUTION: This product is for scientific research use only, and cannot be used for medical or diagnostic procedures on humans or animals, or as food, cosmetics or household products, etc. Without written permission authorisation or approval, this product may not be manufactured, licensed for sale, sold or imported or exported, or use all relevant patents and related trademarks of the product.
Frequently Asked Questions:

1. No amplification product or little product amount
   Check the primer design, confirm the purity and concentration of the primer.
   ② Repeat the experiment to make sure that the reaction system is prepared and the reaction procedure is set correctly.
   ③Increase the number of cycles
   ③Increase the number of cycles ④Increase the concentration of Mg2+ ⑤Optimise the annealing temperature
   ⑤ Optimise the annealing temperature
   (v) Optimise annealing temperature (vi) Try different dosages of blood
2. The product shows high molecular weight dispersed bands.
   The product shows high molecular weight dispersed bands. 1) Centrifuge the supernatant before electrophoresis of the amplified sample.
   (ii) Extension time should not exceed 60 sec/kb.
   (iii) Optimise annealing temperature
   (iii) Optimise annealing temperature (iv) Experiment with different blood volumes (v) Reduce the total number of cycles
   ⑤ Reduce the total number of cycles
   ⑤ Reduce the total number of cycles ⑥ Reduce the primer concentration
3. The amplification product spreads in the gel or disperses in the form of sheet-like bands.
   ①Centrifuge the amplified samples before electrophoresis.
   ②Increase the annealing temperature
   ③Try different amounts of blood
   ④Lower the primer concentration
   ⑤ Reduce the total number of cycles
   ⑥ Redesign the primers