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Revision as of 13:22, 20 October 2010 (view source) Needks (Talk | contribs) (New page: ===Digestion=== # Digestion of PCR products and pSB1A2 ## PyodA-mAAA : EcoRI and SpeI ## PzntA-RFP : XbaI and PstI ## pSB1A3 : EcoRI and PstI <br> {| style="border: 1px solid black;" |- |'...) ← Older edit		Latest revision as of 13:43, 20 October 2010 (view source) Needks (Talk | contribs)
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	===Digestion===		===Digestion===
	# Digestion of PCR products and pSB1A2		# Digestion of PCR products and pSB1A2
-	## PyodA-mAAA : EcoRI and SpeI	+	## PyodA-mAAA : ''Eco''RI and ''Spe''I
-	## PzntA-RFP : XbaI and PstI	+	## PzntA-RFP : ''Xba''I and ''Pst''I
-	## pSB1A3 : EcoRI and PstI	+	## pSB1A3 : ''Eco''RI and ''Pst''I
	<br>		<br>
	{| style="border: 1px solid black;"		{| style="border: 1px solid black;"

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Latest revision as of 13:43, 20 October 2010

Digestion

1. Digestion of PCR products and pSB1A2
   1. PyodA-mAAA : EcoRI and SpeI
   2. PzntA-RFP : XbaI and PstI
   3. pSB1A3 : EcoRI and PstI

Reagent	Volume (μL)
DNA (about 30ng/μL)	30
10x NEB buffer 2	5
BSA (10mg/mL)	0.5
Appropriate 1st and 2nd restriction enzymes	2 (each 1)
D.W.	12.5 / total = 50
Completely digestion at 37˚C for 2 hours (at least) and stop at 80˚C for 20min

1. Confirmation of digested products by agarose gel electrophoresis (Figure 4)
2. Quantification of DNA concentration by NanoDrop