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For 11 individuals (4 of B1 and 7 of B2), PCR reactions were purified using agarose gel extraction kits (Qiagen), and sent for direct sequencing using an AB1 3730XL (Functional Biosciences, Inc., Madison, WI).
PCR reactions with amplicons of expected mass were either purified using the PCR Micro Kit (Invitrogen Australia, Victoria, Australia) and sent for direct sequencing or cloned using the TA-TOPO for Sequencing (pCR-4) cloning kit (Invitrogen Australia, Victoria, Australia), pGEM-T Easy (Promega) or pDrive (Qiagen) according to the manufacturer's instructions.
PCR products were visualized on 1.5% agarose gel using SYBR-Green (Cambrex Bio Science Rockland Inc .. To verify the identity of the PCR products, bands were eluted, DNA was purified (QIAquick gel purification kit, Qiagen) and sent for direct sequencing at the University of Arizona's sequencing facility.
All amplified products were subsequently sent for direct sequencing (see Duneau et al. 2008 for protocols).
PCR amplicons were cleaned with ExoSap-IT (GE Healthcare) and sent for direct sequencing.
The mutant amplicon was then gel-purified and sent for direct sequencing for confirmation.
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Jackson sent for McGuire.
In addition, patients provided an early-morning sputum sample on the morning following admission; this sample was sent for both direct and concentrated smear microscopy (as described below).
For accessions with polymorphic direct PCR sequencing products, up to 10 plasmid DNAs from independent clones were sent for sequencing, respectively.
I directed Ganz to send for a Pittsburgh criminalist who had served us with discretion in the horrific Primm case, reminding him that nothing must be touched until this Mr. Espy and his black bag of dusts and luminous powders arrived.
And a new Twitter account, for @Dave_is_lost, recently began sending messages; G.M. directed two reporters to follow the account.
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Since I tried Ludwig back in 2017, I have been constantly using it in both editing and translation. Ever since, I suggest it to my translators at ProSciEditing.

Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com