Wednesday, February 11, 2015

2 11 2015 DNA Isolation Part One

Today I'm isolating DNA using a Qiagen 96 well plate kit received in October 2014. I'm doing this because samples from the outplant oyster were of considerably poor quality for RAD-Sequencing. Now I'm process samples that were collected last September and October from all three populations at Oyster Bay, Manchester, and Fidalgo Bay. I think these samples are of better quality because of the short time between pulling them from the water and processing the tissues (24-48 hours).

Sample Date:
Oyster Bay    9/19/2014
Fidalgo Bay 10/17/2014
Manchester  10/24/2014

Sample Layout in 96 Well Plate

A1N1-4 11N1-4 21N1-4 31N1-4 41N1-4 51N1-4 61N1-4 71N1-4 81N1-4 111N1-4 101H1-4 11H1-4 2
B1H1-4 31H1-4 41H1-4 51H1-4 61H1-4 71H1-4 81H1-4 91H1-4 101S1-4 11S1-4 21S1-4 31S1-4 4
C1S1-4 51S1-4 61S1-4 71S1-4 81S1-4 91S1-4 102N1-4 12N1-4 22N1-4 32N1-4 42N1-4 52N1-4 6
D2N1-4 72N1-4 82N1-4 92N1-4 102H1-4 12H1-4 22H1-4 32H1-4 42H1-4 52H1-4 62H1-4 72H1-4 8
E2H1-4 92H1-4 102S1-4 12S1-4 22S1-4 32S1-4 42S1-4 52S1-4 62S1-4 72S1-4 82S1-4 92S1-4 10
F4N1-4 14N1-4 24N1-4 34N1-4 44N1-4 54N1-4 64N1-4 74N1-4 84N1-4 94N1-4 104H1-4 14H1-4 2
G4H1-4 34H1-4 44H1-4 54H1-4 64H1-4 74H1-4 84H1-4 94H1-4 104S1-4 14S1-4 24S1-4 34S1-4 4
H4S1-4 54S1-4 64S1-4 74S1-4 84S1-4 94S1-4 10ControlControlControlControlControlControl

I carefully placed each sample in the corresponding microcollection tube by removing the tissue from the original 1.5 ml sample tubes with flame sterilized forceps and gently sliding it into the collection tube. 

Then I created the working buffer by 2 ml proteinase K to 17.5 ml ATL buffer from the Qiagen kit into a 50 ml falcon tube. Then briefly vortexed the mixture and poured it into a channel pipetter liquid vessel. 

Using the 12 Channel Pipetter I pipetted 200 ul of the working buffer into each tube. 

I mixed the solution using inversion and then checked to see if all samples were submerged in the working lysis buffer. 

For samples that were not in the buffer I used a clean 1000 ul pipette tip to gently slide the sample into the buffer.

All samples were fully sumberged in the mixture and are now incubating at 56 C in FTR 228. 

No comments:

Post a Comment