Megabase DNA Extraction from Animal Blood
Collect 3mL animal's blood in 5 mL EDTA anticoagulative tube, and store at 4 °C. 
Mix blood by gently rocking for 10min at room temperature to ensure a homogenous WBC distribution.
Transfer 3ml blood to 15ml screw cap tube containing 9ml RBC lysis solution (3 volumes), and mix by gently inverting 10x.
Incubate 5 min at room temperature.
Gently mix by inverting at least once during incubation.
Spin 5 min at 2000xg at 4⁰C and carefully pipet out supernatant while not disturbing the pellet.
Resuspend cell pellet in 3ml PBS (1 blood volume): 
Add 9ml rbc lysis sol (3 volumes), invert to mix and incubate 5 min at room temperature.
Spin cell suspension for 5 min at 2000xg at 4⁰C and carefully pipet out supernatant while not disturbing the pellet.
Remove the last drop of liquid by tilting the tube and removing liquid with pipet tip.
Resuspend cell pellet in 580ul cell suspension buffer (Bio Rad plug lysis kit) until homogenous suspension.
Transfer 375ul to a new microfuge tube and label (400).
Transfer 187.5ul to another tube labeled (200); add 187.5ul cell suspension buffer, and pipet mix gently 5x.
Keep both tubes on ice until ready to embed in agarose (section II).
Melt 2% agarose (Bio Rad kit) by immersing in microwave-boiled water for 10-15min until agarose is completely melted.
Equilibrate both tubes from step 11 section I in 43°C water bath/heat block for at least 5 min before use.
After 5 min, prepare 5 plugs from one tube by performing the next steps rapidly to avoid solidification of the cell-agarose mixture before pipeting into plug mold: 
Place plug cast on ice for at least 45min or until agarose has solidified.
Alternatively, can place plug cast on inverted metal microfuge ice block on ice for 45min, to avoid potential contact of agarose with ice. 
Wash away hemoglobin and cellular material with Proteinase K and Wash Buffer.
