PCM31
True chip expansion Hi silicon PCR mix

PCM80
Universal FastStart Probe Mixture（UNG）

PCM31 Comparison of Amplification Effects of Different High Fidelity Mixes – Different Species, Different Models

① Brand A ② Brand B ③ TZ ④ PCM31

Newly designed model primers:
Human primers: H5k, H5k high GC, H10k
Mouse Primer: M5k, M5k, High GC.M10k

	Temperature	Time	Loop
Pre denaturation	98℃	30s	1×
Denaturation	98℃	10s	35 cycles
Annealing	H/M/O:60℃	20s
	cDNA- 2.8k:56°C
	cDNA- 4.5k:56°C
Extend	72℃	30s/kb
Final extension	72℃	5min	1×
Low temperature	8℃	Hold	1×

Newly designed model primers
Rice Primers: 05k, o5k – High GC, 010k
cDNA Primers: C-2.8k, C-4.5k

PCM80 Anti inhibition Performance Test: Application of Inhibitors (Ethanol, Blood, EGTA) Test Reagent for Anti inhibition Ability

Comparison of UNG Anti Inhibition CT Values
	3	15	16
UNG	33.54	34.04	34.27
Blood -0.2%	↓1.39 32.15	↓1.41 32.63	↓1.27 32.99
Ethanol -3%	↓1.05 32.49	↓1.99 32.05	↓1.50 32.76
EGTA-2mM	33.38	33.90	34.23

There was no significant difference in CT values when adding 2mMEGTA as the final concentration. However, when adding 0.2% ASFV pig blood or 3% ethanol as the final concentration, the CT values advanced by 1-2 compared to 2xmix. Adding blood signals resulted in varying degrees of reduction (25% -28%).

PCM80 Digestive Ability Test

Application of human genome exploration and 2xmix (UNG) for one round of amplification (20cycles), followed by ten fold gradient dilution of one round of amplification products. The amplification products diluted by 103, 104, 105, 106, and 107 times were selected and amplified using no anti fouling system reaction solution and anti fouling system reaction solution, respectively. The CT differences at the same template concentration were compared.

Comparison of human genome template amplification:

Comparison of amplification of products containing dU in one round:

Conclusion: By amplifying 1ng/uL gDNA, PCM80 can be performed about 2 times earlier than UNG CT without anti fouling system; After amplifying the product containing dU in one round, PCM80 showed a significant CT lag effect compared to UNG without anti fouling system (CT difference greater than 10). When the product was diluted 106 times in one round, PCM80 did not amplify, indicating a direct relationship with the UNG system and strong digestive ability.