User:Jessica Karen Wong/Progress

T9002

Output Measurement Kit (for inverters, terminators, etc) Using GFP

<BBpart> T9002</BBpart>

• F2620 has Mfe/BB prefix tails
• Got only 1 right size colony from 3K3 ligation/transformation
• Waiting on seq verification
• E0240 has been PCR'ed with tails, looks the right size
• Digesting E0240 S/N to ligate into F2620

E0240

Promoter Tester with GFP

<BBpart> E0240</BBpart>

• Part is built
• PCRed with Mfe1 and Spe1 tails
• Successfully ligated into 3K3 - Sequence verifies that it's scarred
• Inserted a BB site between plasmid and RBS
• Made both E/S and E/X versions
• Has a 1bp mutation around 700
• Did mutagenesis and got a good number of colonies
• Sent in right size colonies for seq

I2057

Promoter tester with RFP

<BBpart> I2057</BBpart>

• Part is built (sequence verified)
• PCR-ed with Mfe1 and Nsi1
• Ligated in 3K3 (seq verified scarred)
• Inserted a BB site b/t plasmid and RBS
• Created both E/X and E/S versions
• E/S sealed w/ Not1 is seq verified 267-end, waiting on redo for beginning
• E/X sealed w/ Not1 is seq verified 1-769, waiting on redo for end

Ligated J116, J100, and R0040 into EX version from PCR - waiting on seq

I2055

RBS tester with GFP

<BBpart> I2055</BBpart>

• Most likely built and scarred
• waiting on redo of seq to make sure middle (bp 95-208) is ok
• Inserted a BB site b/t Promoter and GFP
• Made both E/X and S/X forms
• Sealed both EX and SX with Not1 - waiting on seq
• Inserted B0034 into both EX and SX versions of PCR, B0032 into SX PCR (all SX ligations took place w/ restriction enzymes)

I2056

RBS tester with RFP

<BBpart> I2056</BBpart>

• Formerly called blue

• Part is most likely built and scarred
• Waiting on redo of seq to make sure front is scarred
• Inserted a BB site b/t promoter and RFP
• Created both E/X and S/X
• Attempting to seal both varieties and has same RBS's as I55