deletions | additions       diff --git a/16S rDNA Sequencing and Analysis (Organism Identification).md b/16S rDNA Sequencing and Analysis (Organism Identification).md  index 0703a11..1c54ad2 100644  --- a/16S rDNA Sequencing and Analysis (Organism Identification).md  +++ b/16S rDNA Sequencing and Analysis (Organism Identification).md 
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Centrifuge 1 ml of the overnight culture until the cells form a pellet at the bottom of the tube (about 5 minutes at 10,000 g), pour off the liquid on top (the supernatant) and resuspend the pellet in 100 μl of sterile DNAase-free water. Incubate the samples at 100°C for 10 minutes to help lyse the cells. Use the resulting solution as the template in the PCR reaction below.  ##PCR reaction  This reaction uses the 27F (AGAGTTTGATCMTGGCTCAG) and 1391R (GACGGGCGGTGTGTRCA) primers which amplify a near full-length bacterial (and many archaeal) 16S rRNA gene. Our lab uses standard PCR reagents (Qiagen or Kappa), with an annealing temperature of 54°C 54 °C  and an extension at 72°C 72 °C  of 90 seconds. Do not forget to include positive (any sample containing bacterial genomic DNA that you have successfully amplified before) and negative (_e.g_., replace DNA with water) controls. The full program we use is:  1: 95 °C for 2:00  2: 95 °C for :15  3: 54 °C for :30  4: 72 °C for 1:30  5: Go to 2 (40 times)  6: 72 °C for 3:00  7: 4 °C forever  After PCR is completed, confirm the PCR reaction worked by agarose gel electrophoresis, all controls behaved as expected, and that you have DNA fragments of the correct size (~1350bp).