DNA Quantification Continuation of Lab 4

Qiagen DNAeasy kit. Quantify our yields using a Nanodrop system.

DNeasy Minispin Column Collection tube Eppendorph

Figure 4. Summary of DNA extraction protocol. II.A. DNA Extraction and Purification II. DNA Extraction, Purification and Quantification

1.Use pipet to transfer DNA mixture into DNeasy minispin column 4. Save column. 2. Centrifuge at 8000 rpm for 1 min. 3. Discard liquid and collection tube. STEP 5

1.Use new collection tube 2.Add 500 µl of AW1 buffer 3.Centrifuge 1 minute at 8000 rpm 4.Discard flow-through and collection tube 5.Keep column Flow-through Collection tube STEP 6

1.Use new collection tube 2.Add 500 µl of AW2 buffer 3.Centrifuge 3 minute at rpm 4.Discard flow-through and collection tube 5.Keep column Flow-through Collection tube STEP 7

STEP 8 1.Place Dneasy mini-spin column in 1.5 ml microcentrifuge tube (eppindorph) 2.Snip cap off of column microcentrifuge tube

STEP 9 1.Add 100 µl of AE buffer over disk in column 2.Incubate at room temp for 1 minute 3.Centrifuge 1 minute 8000 rpm 4.DO NOT DISCARD FLOW-THROUGH disk

STEP 10 1.Label with sharpie at top of lid 2.Freeze and save for PCR Lab 10 1.Add another 100 µl of AE buffer over disk in column and repeat procedure below 2.Incubate at room temp for 1 minute 3.Centrifuge 1 minute 8000 rpm 4.DO NOT DISCARD FLOW-THROUGH

Nanodrop spectrometer used to measure DNA concentrations II.B. DNA Quantitation