IIT Madras/20 August 2011

From 2011.igem.org

Lab work planned

1) Agarose Gel of pET41a 1.1)Prepare TAE Buffer, 0.5M EDTA 1.2)Prepare Agarose Gel, Obtain 1kb Ladder L1 - Ladder L2 - pET41a 2) Restriction of pET41a for checking RE Activity 2.1) Cut pET41a at EcoR1, Xba1, Spe1, Pst1 2.2) Run Gel Electrophoresis 3)Transform DH5a with the following parts: GFP - E0040 - Plate 1 - 14K RFP - E1010 - Plate 1 - 18F Single Terminator - B0010 - Plate 1 - 13D Double Terminator - B0015 - Plate 1 - 23L RBS - B0034 - Plate 2 - 2M pCSTAG - K118011 - Plate 2 - 18D 3.1) Restriction with E, P 4) Double Digest pSB1A3, pSB1C3, pSB1K3 5)Miniprep J04450 From Both Plates 5.1) Restrict with E, P 5.2) PCR Purification followed by Test Ligation 5.3) GEL , Elution 6) Puc19 6.1) Obtain 6.2) Miniprep 6.3) Double Digest