1

Round2----- 0 Hrs ----- ------ 3 Hrs ----1 2 3 4 5 6 7 8 1 2 3 4 5 6 7 8

1 2 3 4 5 6 7 81 2 3 4 5 6 7 8

PCR result;-1 pot type: construct No. 2 get non specific product

- 2 Round type; all works!

Transcription result;

1 pot type & 2 Round type; all works, in different amount of RNA

Date: 27 May 2010Exp: RNA game 4

2

I show you sequencing data by using RNA made from R2, construct 1 - 8No.1 = 2782 ( Stephanie’s 3rd)No.2 = 2785 (Hui’s Cross 2)No.3 = 2788 (Chris’s B Cross2)No.4 = 2791 (Jee’s Bulgeless Cross 1)No.5 = 2792 (Jee’s Bulgeless Cross 2)No.6 = 2793 (Jee’s Cross 3)No.7 = 2794 (Adrien’s Bulgeless 1)No.8 = 2795 ((Adrien’s Bulgeless 2)

Abbreviations: NM

Abbreviations;

NM = No modify

ddGTP = use dideoxy-Guanine nucleotide asa

internal ladder

SH = SHAPE reaction

As NM as a background

Construct No.3 = 2788 (Chris’s B Cross2) and No.6 = 2793 (Jee’s Cross 3) seem to be a cleanest construct.

Please look at the SHAPE data, last column of each construct result;

Construct 2 – 7 show the region that corresponds to ‘the loop” [Red arrow], please see the structure image.

3

4

Next thing to do;

1) From the latest result, Chris can assign the sequence from this data.

2) Somehow, my result on ddNTPs should be repeat. Once I finish, I will pass the result to Chris ASAP.

See you all at the meeting!

Regards,

Ann