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October 1st, 2009
For Construct no. 51 (elst_AEA) --> vector with AEA and His-tag on N-term. was cut, isolated from gel and ligated with ELST.
Control restriction of vector with His-tag on N-term. containing EWAND or TetShack showed that none of this inserts was present in vector.
Ligation of construct LL37_ELST_GCN_ELST (front insert) and P1/P2 (front vector) was set.
Vectors containing constructs ELST-ELST-ELST-mNGF, p53, GyrB-p53, foldon-p53, p53-APH, APH-p53, BCR-p53 and p53-APH1 were isolated.
Control restriction was performed on vectors supposedly containing construct ELST-ELST-ELST-mNGF.
Insert without His -tag was ligated in vector.
October 2nd, 2009
Ligation of Construct no. 51 (elst_AEA) was transformed into DH5a.
Transformation of constructs LL37_ELST_GCN_ELST_P1/P2 into DH5a.
Vector containing p53-BCR was isolated from mini-preps.
Control restriction was performed on vectors supposedly containing construct LL37-ELST-GCN-ELST-mNGF.
RADA-mNGF and p53-APH peptides were produced.
October 3rd, 2009
Vector containing LL37-ELST-GCN-ELST-mNGF was isolated from mini-preps.
October 4th, 2009
Isolation and control restriction of constructs LL37_ELST_GCN_ELST_P1/P2.
Western blot was performed to confirm the presence of RADA-mNGF and p53-APH.
Control restriction was made on several clones of vector that is not supposed to contain His-tag. Bands on gel were not visible.
October 5th, 2009
Electrophoresis of PCR product. Isolation from gel.
Constructs LL37_ELST_GCN_ELST_P1/P2 and KSI_DP_P2/33_LL37_ELST_GCN_ELST were sent to be sequenced.
Control restriction was made on several clones of vector that is not supposed to contain His-tag.
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