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Meanwhile, 9 mutants (R18A, L143A, E153A, G182A/F183A, C374A, C376A/G377A, Y393A, V566A and V571A) produced absolute blue colonies (Fig. 2C), and the recombination frequency was zero as compared with wild-type protein (Table 2).
PCR amplification was run with 3.5 μL of cDNA template in 10 μL of reaction mixture containing 5 μL absolute blue qPCR SYBR green ROX mix (Thermo Scientific) and 300 nM primers.
The qPCR reaction (total volume of 20 μl) was prepared by mixing 10 μl of the 2x concentrated absolute blue solution with 1 μM of each kDNA minicircle specific primers JW11 and JW12 (Table 1) and template DNA (2 μl) [ 12].
qPCR reaction mix consisted of 1X Absolute blue QPCR SYBR Green Mix (Thermo Scientific) with 200 nM of forward and reverse primers (GAPDH & Fgf8; Table 1).
Briefly, PCR was performed with the ABsolute blue QPCR SYBR Green ROX mix (Abgene, Thermo Fisher Scientific), using 11 ng of cDNA per RT.
Amplification reactions were performed in 1 × Absolute blue qPCR SYBRGreen fluorescein mix (Thermo scientific) with 5 ng of cDNA template and 300 nM of each primer in a final volume of 15 μL using an iCycler iQ™ thermocycler (Bio-Rad).
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It is the absolute wild blue yonder of Coltrane's scream-music.
I came for a walk and found an absolute true-blue Yorkshireman wandering around in the woods.' It turned out to be Steve Warrillow, the warden who is paid by the factory to manage the reserve on a full-time basis.
"I woke up the morning after the election and saw that Brighton and Hove was now just a tiny red and green island in an absolute sea of blue.
The absolute number of blue cells was counted out of 2000 cells.
Before the Light in the Attic reissues, fans such as Devendra Banhart and Nick Cave ("she is my absolute favourite female blues singer") had to dig deep for her albums.
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