2X PCR Mix

2 X High Fiderity Polymerase PCR Mix from Thermofisher For Cloning

	for 100 ul
H2O	48 ul
5 X  HiFi PCR Buffer	40 ul
10 mM dNTP	4 ul
5 Units/ul of Superfi II from Thermofisher	4 ul

Mix, Spin briefly, Store at -20 oC

Use 5 ul for PCR, final volume is 10 ul

For 10 ul PCR

F primer: 0.5 ul

R Primer: 0.5 ul

templete, dip it; genomic DNA, 1 ul

2 X PCR mix: 5 ul

water: to 10 ul

2 X High Fiderity Polymerase PCR Mix For Cloning

		for 1 X 25 ul reaction	for 10X 25 ul reactions
10 mM dNTP		1 ul	11 ul
5 Units/ul of Q5, or Kfu, or Pfu or Kapa or Phusion		0.5 ul	5.5 ul
5 X  Q5 PCR Buffer		5 ul	55 ul
or 10 X  HiFi PCR Buffer		or 2.5 ul	or 27.5 ul
H2O		6 ul if 5X buffer is used 8.5 ul if 10X Buffer is used	203.5 ul if 5X Buffer is used 231 ul if 10 X Buffer is used

2X Taq PCR Mix for Analysis

		for 1 X 25 ul reaction	for 10X 25 ul reactions
10 mM dNTP		1 ul	11 ul
5 Units/ul of Taq		0.5 ul	5.5 ul
10 X Taq PCR Buffer		2.5 ul	or 27.5 ul
H2O		8.5 ul	231 ul

PT Mix (Primers + Template)

		for 1 X 25 ul reaction	for 10X 25 ul reactions
10 uM Forward Primer		1 ul	11 ul
10 uM Reverse Primer		1 ul	11 ul
Optional: Template (miniprep DNA, or E coli culture)		0.1~0.5 ul	0.1~0.5 ul
H2O		10.5 ul	103 ul

On ice, aliquote 12.5 ul of 2 X PCR Mix to each PCR tube, add 12.5 ul PT Mix Add 0.1 to 0.5 ul of  template DNA sample to each tube if the PT Mix does not contain template. For example, E coli culture from colonies.