Revision as of 04:38, 4 January 2007

SAGA deletion Module

need quiz for this day to start lab
need FY2068 ON then subcultured flask growing for lab
need YPD plates, and -ura and a few -trp
aliquots for each pair of students: 1 ml ON culture, 1 ml of log phase, 5 ml sterile H20 in conical tube

must wash and reprobe with dendrimers between this lab and next

YPD
Buffer Y1 for RNA isolation: 1M sorbitol (9.1g/50ml), 0.1M EDTA (10 ml of 0.5M/50 ml), Filter sterilize, leave at RT
Zymolyase Stock: MP Biomed #32093 = 100,000U/g so 10 mg/100 ul Y1 gives 100U/10 ul.
Working spheroplast solution (per sample): 1 ml Y1 +10 ul Zymolylase stock + 1 ul BME.
RLT+BME for RNA prep; 1 ml RLT from quiagen + 10 ul BME. Use 350 ul/sample
0.5M NaOH/0.5M EDTA: 0.1g NaOH to 5 ml 0.5M EDTA (check this calculation).
6X SSC/0.005% Triton: 150 ml 20X SSC, 25 ul TritonX-100, 350 ml H20 (how many bottles needed?)
2XSSC/0.0016% Triton: dilute 6X 1:3
0.2X SSC/0.00016% Triton: dilute 6X 1:30

@@ Line 7: / Line 7: @@
 #need FY2068 ON then subcultured flask growing for lab
 #need YPD plates, and -ura and a few -trp
+#aliquots for each pair of students: 1 ml ON culture, 1 ml of log phase, 5 ml sterile H20 in conical tube
 ===Notes:M3D3===
 ===Notes:M3D4===