User:Karmella Haynes/Notebook/BioBrick cloning/2013/03/06

Current revision

Karmella's BioBrick Cloning

Colony PCR

Streaked single colonies on gridded plates yesterday
Used bit of streak for PCR today
All parts are on pSB1A3-based vectors (http://partsregistry.org/Part:pSB1A3)
- Use primers VF2 and VR to amplify inserts

Reagent	Volume	Expected: Primers add ~250bp 1-3. K084007 (835 bp) = 1085 4-6. R0071 (53 bp) = 303 7. K081016 (735 bp) = 985	10 μL/lane; 1% agarose; Ladder
Bacteria from tip	---
10 μM VF2 primer	1.0
10 μM VR primer	1.0
2x GoTaq Green	12.5
dH₂O	10.5
	25.0

PCR

@@ Line 16: / Line 16: @@
 * Used bit of streak for PCR today
 * All parts are on pSB1A3-based vectors (http://partsregistry.org/Part:pSB1A3)
+** Use primers VF2 and VR to amplify inserts
 {| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table -->
@@ Line 21: / Line 22: @@
 | bgcolor=#cfcfcf | Reagent
 | bgcolor=#cfcfcf | Volume
-| rowspan="7" | <u>Expected:</u><br>Primers add ~250bp<br>1-3. BB 1 (# bp) = ##<br>4-6. R007.. (# bp) = ##<br>7. K.. (# bp) = ##
+| rowspan="7" | <u>Expected:</u><br>Primers add ~250bp<br>1-3. K084007 (835 bp) = 1085<br>4-6. R0071 (53 bp) = 303<br>7. K081016 (735 bp) = 985
 | rowspan="7" | [[Image:KAH030713_gel1.jpg|300px|PCR gel]]<br>10 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
 |-