analyze pcr please. picture condition is not a good due to machine disorder Well

Question

analyze pcr please. picture condition is not a good due to machine disorder

Well #

Label

Row 1 of Gel (S11F1 and S11R1)

Label

Row 2 of Gel (HHP1 primers)

1

Ladder

Ladder

2

1-4-a

cDNA 1 WT + F/R

2-4-a

cDNA 1 H2O2+ F/R

3

1-4-b

cDNA 1 WT + F/R

2-4-b

cDNA 1 WT +F/R

4

1-4-c

cDNA 2 WT + F/R

2-4-c

cDNA 2 H2O2+ F/R

5

1-4-d

cDNA 2 WT + F/R

2-4-d

cDNA 2 WT + F/R

6

1-4-e

cDNA 1 H2O2 + F/R

2-4-e

cDNA 1 WT + no primers (neg control)

7

1-4-f

cDNA 1 H2O2 + F/R

2-4-f

cDNA 2 WT + no primers (neg control)

8

1-4-g

cDNA 2 H2O2 + F/R

9

1-4-h

cDNA 2 H2O2 + F/R

Tube label

Tube contents

1-4-a

2µl cDNA1-9 + 2.5µl S11F1 + 2.5µl S11R1 + AccuPower + 13µl H2O

1-4-b

2µl cDNA1-9 + 2.5µl S11F1 + 2.5µl S11R1 + AccuPower + 13µl H2O

1-4-c

2µl cDNA1-10 + 2.5µl S11F1 + 2.5µl S11R1 + AccuPower + 13µl H2O

1-4-d

2µl cDNA1-10 + 2.5µl S11F1 + 2.5µl S11R1 + AccuPower + 13µl H2O

1-4-e

2µl cDNA1-9 + AccuPower + 18µl H2O

1-4-f

2µl cDNA2-8 + 2.5µl S11F1 + 2.5µl S11R1 + AccuPower + 13µl H2O

1-4-g

2µl cDNA2-8 + 2.5µl S11F1 + 2.5µl S11R1 + AccuPower + 13µl H2O

1-4-h

2µl cDNA2-7 + 2.5µl S11F1 + 2.5µl S11R1 + AccuPower + 13µl H2O

2-4-a

2µl cDNA2-7 + 2.5µl S11F1 + 2.5µl S11R1 + AccuPower + 13µl H2O

2-4-b

2µl cDNA2-7 + AccuPower + 18µl H2O

2-4-c

2µl cDNA1-10 + 2.5µl FHHP1 + 2.5µl RHHP1 + AccuPower + 13µl H2O

2-4-d

2µl cDNA1-9 + 2.5µl FHHP1 + 2.5µl RHHP1 + AccuPower + 13µl H2O

2-4-e

2µl cDNA2-8 + 2.5µl FHHP1 + 2.5µl RHHP1 + AccuPower + 13µl H2O

2-4-f

2µl cDNA2-7 + 2.5µl FHHP1 + 2.5µl RHHP1 + AccuPower + 13µl H2O

Accupower Quickstart PCR

WT cDNA1 = cDNA 1-9

H2O2 cDNA1 = cDNA 1-10

WT cDNA2 = cDNA2-7

H2O2 cDNA2 = cDNA2-8

Lysis times: 1-9, 1-10 11:15am-2:11pm

          2-7, 2-8 12:15pm-2:46pm    

PCR ran at 56C

Well #

Label

Row 1 of Gel (S11F1 and S11R1)

Label

Row 2 of Gel (HHP1 primers)

1

Ladder

Ladder

2

1-4-a

cDNA 1 WT + F/R

2-4-a

cDNA 1 H2O2+ F/R

3

1-4-b

cDNA 1 WT + F/R

2-4-b

cDNA 1 WT +F/R

4

1-4-c

cDNA 2 WT + F/R

2-4-c

cDNA 2 H2O2+ F/R

5

1-4-d

cDNA 2 WT + F/R

2-4-d

cDNA 2 WT + F/R

6

1-4-e

cDNA 1 H2O2 + F/R

2-4-e

cDNA 1 WT + no primers (neg control)

7

1-4-f

cDNA 1 H2O2 + F/R

2-4-f

cDNA 2 WT + no primers (neg control)

8

1-4-g

cDNA 2 H2O2 + F/R

9

1-4-h

cDNA 2 H2O2 + F/R

Explanation / Answer

Figure 1

When we use primers- (S11F1 and S11R1) we got the amplification only in the case of cDNA 1 WT.

This shows that the gene, that we are looking, is only expressed in sample 1 which we labelled as cDNA 1. When the cells were treated with H2O2, the band which we were getting in the wild-type 1 was not expressed when the same cells were treated with H2O2.

Figure 2 - HHP1 primers

When the cells were treated with H2O2 we got the band in cDNA 1 which was faint or the band we got in the gel was due to leakage during the sample loading in the well in wild-type cells. In case of sample 2 or cDNA 2, there was the very little expression in both, H2O2 and wildtype cells.

These results show that gene for which primers S11F1 and S11R1 was used only expressed in wild-type of sample 1.

These results show that gene for which primers HHP1 primers was used only expressed in H2O2 treated sample 1 and very less expression in wild-type and H2O2 treated sample 2.

Write me back if you still have doubt. Don't forget to hit thumbs up button

Related Questions

Navigate

• Browse (All)
• Browse A
• Subjects

---

---

Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.