Page 18 - 2022-23 BIOFACT Catalogue

PCR Description

PCR (Polymerase Chain Reaction) is a principle where target regions of genomic DNA or cDNA are selected and the desired ones are ampli-
ed to make multiple copies. This technique undergoes the repetitive stage of the following PCR system. Then, in an in vitro setting, DNA
is ampli ed by more than 10 times minimally within several hours, which corresponds to its methods. Each PCR enzyme of the BIOFACT
5
is expressed in multiple amounts following the recombination of genes coding high-temperature DNA polymerase in E. coli. Thus, it is a
thermal resistant DNA polymerase that has been puri ed at a higher degree of purity.

Target gene Primer PCR Result analysis
selection design reaction (For electrophoresis
system)

PCR Workflow

PCR System (Thermal Cycling) Primer Design Guide
cycle #n It is recommended to design the primer as follows:
Chemistry selection Primer and 1. Length : 17 ~ 28 mer (Tm = 55 ~ 80 ℃) Data
qPCR
(DNA-binding Dyes probe design 2. Base composition : 50 - 60% (G+C) analysis
optimization
or Probe type) A
B 3. Setting of Tm value of forward primer to be similar to Tm value
C
D of reverse primer
E
4. Region to avoid : 1) G + C rich region
F
2) Repeated region and secondary structure
5’ 3’
3’ 5’
1’ Design Rule (= The Wallace rule)
1 temperature 1 1 1 *Annealing Temperature =Tm – (4 ~ 6 ℃)
Tm (℃) = 2(A+T) + 4(C+G)
3 3 3 3
2 2 2 2 (A+T) = Sum of A and T residues in the primer
0 cycle #1 cycle #2 cycle #3 cycle #4 (C+G) = Sum of C and G residues in the primer
PCR Additives ; 5X Band Helper™
1. Pre-denaturation T
2. Denaturation step ① 5X Band Helper™ is an innovative PCR additive which is highly suit-
Cycling able for use in all PCR enzymes of BIOFACT. It enables e cient am-
3. Primer annealing (Annealing step) ②
(20~40 cycle) pli cation and improved results can be obtained by adjusting of 5X
4. Extension step ③ Band Helper™ in the following cases.
5. Final extension step - Di cult templates which are GC-rich or have a high degree of
secondary structure
- Presence of non-speci c bands or products
* It is recommended to use a minimum amount of 5X Band Helper™ to avoid
the mutation in case of PCR enzymes that have delity features.
BIOFACT Product
F-Star Taq DNA Polymerase (p. 78), Thumb Taq DNA Polymerase (p. 80), Taq DNA Polymerase (p. 82)
Lamp Taq DNA Polymerase (p. 88), Pfu DNA Polymerase (p. 92), Lamp Pfu DNA Polymerase (p. 94)

18 BIOFACT CATALOGUE