Short protocols, superior performance

  1. Buccal swab (0.5 mm punch)
  2. Hair (1 mm)
  3. Tooth (1 mm piece)
  4. Nail (1x2 mm)
  5. Saliva (0.5 µl)
  6. Amniotic fluid (1.0 µl)
Consistent results from various human samples with Direct PCR. Different samples were placed directly into 20 µl PCR reactions. A 237 bp DNA fragment was successfully amplified using the control primers included in the kit. Reactions were run in Thermo Scientific UTW reaction vessels using Thermo Scientific Piko Thermal Cycler. + and – denote control reactions with and without purified DNA.
 
 
kb
Total cycling time (min)
Direct PCR Hot start Taq
0.5 30 84
1.0 30 84
1.5 35 99
3.8 62 202*

*no product

Significant time savings and superior performance in comparison to conventional PCR from purified DNA. Four DNA fragments (0.5–3.8 kb) were amplified directly from buccal swabs using the dilution protocol. For the comparison reactions, DNA was first purified using a DNA purification kit from a major supplier. The same DNA fragments were then amplified using a hot start Taq polymerase following the manufacturer's instructions. Thermo Scientific Phusion Human Specimen Direct PCR Kit delivered greater yields and was also able to reliably amplify the longer fragments. Comparison of total cycling times of both methods shows that the Phusion Human Specimen Direct PCR Kit is 2–3 times faster allowing significant time savings in PCR.