From 2007.igem.org

Revision as of 05:05, 27 October 2007 (view source) Silvapy (Talk | contribs) ← Older edit		Revision as of 05:16, 27 October 2007 (view source) Silvapy (Talk | contribs) Newer edit →
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-	'''Materials'''	+	== '''Materials''' ==
-	'''Methods'''	+	== '''Methods''' ==
-		+	'''Primer Design'''
-	PCR	+
-	Dephosphorylation	+
-	Sequencing	+
	Biobricking of bch-gene cluster and crtN gene.		Biobricking of bch-gene cluster and crtN gene.
Line 35:		Line 32:
	bch B      3'ttaGGATCCccgttcgccttggtttgacttact5'		bch B      3'ttaGGATCCccgttcgccttggtttgacttact5'
-	'''Results'''
-	'''Discussion'''	+	PCR
		+	PCR of bch-? genes using - 20ng/ul pHM6
		+	1ul pHM6
		+	1ul dNTP
		+	5ul primer mix
		+	0.5ul enzyme-phusion polymerase
		+	32.5ul H20
		+	50ul total
		+
		+	'''Dephosphorylation'''
		+
		+	== '''Results''' ==
		+
		+	'''Sequencing'''
		+
		+	== '''Discussion''' ==

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Revision as of 05:16, 27 October 2007

Contents 1 Materials 2 Methods 3 Results 4 Discussion

Materials

Methods

Primer Design

Biobricking of bch-gene cluster and crtN gene.

Gene:

bchB

bchE

bchI

bchL

bchM

bchN

[[crtNgene]]

crtN

PCR of crtN on Heliobacillus mobilis G/C content on annealing region

crtN-F 40% BglII CrtN-R 50% BamHI and XhoI

Primers

crtN-F 3’ccaaaAGATCTgtgaaacatacagcaaaaaacctgggt5’

crtN-R 5’gctagCTCGAGttaGGATCCtcagtaactggctgacaagcct3’

bch B 3'ttaGGATCCccgttcgccttggtttgacttact5'

PCR

  PCR of bch-? genes using - 20ng/ul pHM6
  1ul pHM6 
  1ul dNTP
  5ul primer mix 
  0.5ul enzyme-phusion polymerase
  32.5ul H20
  50ul total
   

Dephosphorylation

Results

Sequencing

Discussion